triplicate steponeplus Thermo Fisher Search Results


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  • 90
    Thermo Fisher steponeplus instrument
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Instrument, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 30994 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    steponeplus instrument - by Bioz Stars, 2020-04
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    94
    Thermo Fisher steponeplus cycler
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Cycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 297 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    steponeplus cycler - by Bioz Stars, 2020-04
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    Thermo Fisher steponeplus thermocycler
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Thermocycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 762 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher steponeplus
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3786 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher steponeplus system
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4108 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi steponeplus thermocycler
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Abi Steponeplus Thermocycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    abi steponeplus thermocycler - by Bioz Stars, 2020-04
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    96
    Thermo Fisher steponeplus platform
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Platform, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher steponeplus sequence detection system
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 325 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi steponeplus
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Abi Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 353 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi steponeplus system
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Abi Steponeplus System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher applied biosystem steponeplus
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Applied Biosystem Steponeplus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher steponeplus machine
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Machine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 505 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    steponeplus machine - by Bioz Stars, 2020-04
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    Thermo Fisher steponeplus thermal cycler
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Steponeplus Thermal Cycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 398 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qpcr steponeplus life technologies
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Qpcr Steponeplus Life Technologies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher applied biosystems steponplus instrument
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Applied Biosystems Steponplus Instrument, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi steponeplus apparatus
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Abi Steponeplus Apparatus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher applied biosystems steponeplus device
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Applied Biosystems Steponeplus Device, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher power sybr green detection
    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Power Sybr Green Detection, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
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    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
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    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
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    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e <t>qRT-PCR</t> showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX
    Stepone Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14067 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e qRT-PCR showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX

    Journal: Stem Cell Research & Therapy

    Article Title: Targeted genome engineering in human induced pluripotent stem cells from patients with hemophilia B using the CRISPR-Cas9 system

    doi: 10.1186/s13287-018-0839-8

    Figure Lengend Snippet: CRISPR-Cas9 and insertion of F9 cDNA into AAVS1 locus of HEK293T cells. a Schematic of CRISPR-Cas9. b About 80% of cells were GFP-positive 24 h after transfection in the GFP group. c Primers F1, R1 were used to detect the full fragment of insertion. In the 293 T-WT group, only a 468-bp fragment could be seen (yellow arrow); in the 293 T-insertion group, both 468-bp and 4.9-kb fragments could be seen (yellow arrow). d. Primers F2, R2 used to detect 5′ junction point of insertion. In the 293 T-WT group, nothing could be seen; in the 293 T-insertion group, a 1.3-kb fragment could be seen (yellow arrow). e qRT-PCR showed F9 expression of the 293 T-insertion group was extremely higher than that in the 293 T-WT group. AAVS1-EF1α-puromycin empty plasmid did not produce any hFIX expression (n = 3 independent experiments for each sample). f Immunofluorescence staining showed hFIX expression in the 293 T-insertion group. All scale bars represent 100 μm. CRISPR-Cas9 clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9, WT wildtype, GAPDH glyceraldehyde 3-phosphate dehydrogenase, DAPI 4′,6-diamidino-2-phenylindole, HFIX human factor IX

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed in triplicate, using a Step-One-Plus Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: CRISPR, Transfection, Quantitative RT-PCR, Expressing, Plasmid Preparation, Immunofluorescence, Staining

    Differentiation of iPSCs into hepatocytes and characterization of hepatocytic functions. a Schematic showing stepwise protocol for producing hepatocytes from iPSCs. b Immunofluorescence staining showed expression of SOX17, FOXA2, and GATA4 on day 5, HNF4α on day 10, AFP on day 15, and ALB on day 20. c Flow cytometry analysis showed expression of AFP and ALB > 90%. d qRT-PCR showed relative expression of HNF4α, AFP, ALB, TDO2, and TAT relative to GAPDH (n = 3 independent experiments for each sample). e Expression of pluripotent markers (OCT4, SOX2, and NANOG) and hepatic markers (AFP, ALB, HNF4α, TAT, TDO2, and CYP3A4) between iPSC-insertion and iPSC-insertion-Heps compared by qRT-PCR (n = 3 independent experiments for each sample). After differentiation, expression of pluripotent markers decreased, while expression of hepatic markers increased. All scale bars represent 100 μm. BMP bone morphogenetic protein, bFGF basic fibroblast growth factor, iPSC induced pluripotent cell, HGF, hepatocyte growth factor, DAPI 4′,6-diamidino-2-phenylindole, AFP alpha-fetoprotein, ALB albumin, GAPDH glyceraldehyde 3-phosphate dehydrogenase

    Journal: Stem Cell Research & Therapy

    Article Title: Targeted genome engineering in human induced pluripotent stem cells from patients with hemophilia B using the CRISPR-Cas9 system

    doi: 10.1186/s13287-018-0839-8

    Figure Lengend Snippet: Differentiation of iPSCs into hepatocytes and characterization of hepatocytic functions. a Schematic showing stepwise protocol for producing hepatocytes from iPSCs. b Immunofluorescence staining showed expression of SOX17, FOXA2, and GATA4 on day 5, HNF4α on day 10, AFP on day 15, and ALB on day 20. c Flow cytometry analysis showed expression of AFP and ALB > 90%. d qRT-PCR showed relative expression of HNF4α, AFP, ALB, TDO2, and TAT relative to GAPDH (n = 3 independent experiments for each sample). e Expression of pluripotent markers (OCT4, SOX2, and NANOG) and hepatic markers (AFP, ALB, HNF4α, TAT, TDO2, and CYP3A4) between iPSC-insertion and iPSC-insertion-Heps compared by qRT-PCR (n = 3 independent experiments for each sample). After differentiation, expression of pluripotent markers decreased, while expression of hepatic markers increased. All scale bars represent 100 μm. BMP bone morphogenetic protein, bFGF basic fibroblast growth factor, iPSC induced pluripotent cell, HGF, hepatocyte growth factor, DAPI 4′,6-diamidino-2-phenylindole, AFP alpha-fetoprotein, ALB albumin, GAPDH glyceraldehyde 3-phosphate dehydrogenase

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed in triplicate, using a Step-One-Plus Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Immunofluorescence, Staining, Expressing, Flow Cytometry, Cytometry, Quantitative RT-PCR

    Generation and characterization of patient-specific iPSCs from PBMNCs. a Schematic of patient-specific iPSC generation from PBMNCs. b Small iPSC-like colonies appeared 7 days after electroporation. At 14 days after electroporation, iPSC colonies grew big enough for picking out. After being picked out, iPSCs were cultured on Matrigel-coated plates without feeder. c Karyotype of iPSCs was normal. d qRT-PCR analysis showed expression of OCT4, SOX2, and NANOG of iPSCs ( n = 3 independent experiments for each sample). PBMNCs from patient used as negative control, H1 embryonic stem cells used as positive control. e Immunofluorescence staining showed expression of TRA-1-60, SSEA4, OCT4, and NANOG. f Sections of teratomas stained with H E (endoderm: bronchus; mesoderm: cartilage; ectoderm: epidermis). All scale bars represent 100 μm. PBMNC peripheral blood mononuclear cell, iPSC induced pluripotent cell, D day, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HB hemophilia B

    Journal: Stem Cell Research & Therapy

    Article Title: Targeted genome engineering in human induced pluripotent stem cells from patients with hemophilia B using the CRISPR-Cas9 system

    doi: 10.1186/s13287-018-0839-8

    Figure Lengend Snippet: Generation and characterization of patient-specific iPSCs from PBMNCs. a Schematic of patient-specific iPSC generation from PBMNCs. b Small iPSC-like colonies appeared 7 days after electroporation. At 14 days after electroporation, iPSC colonies grew big enough for picking out. After being picked out, iPSCs were cultured on Matrigel-coated plates without feeder. c Karyotype of iPSCs was normal. d qRT-PCR analysis showed expression of OCT4, SOX2, and NANOG of iPSCs ( n = 3 independent experiments for each sample). PBMNCs from patient used as negative control, H1 embryonic stem cells used as positive control. e Immunofluorescence staining showed expression of TRA-1-60, SSEA4, OCT4, and NANOG. f Sections of teratomas stained with H E (endoderm: bronchus; mesoderm: cartilage; ectoderm: epidermis). All scale bars represent 100 μm. PBMNC peripheral blood mononuclear cell, iPSC induced pluripotent cell, D day, GAPDH glyceraldehyde 3-phosphate dehydrogenase, HB hemophilia B

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed in triplicate, using a Step-One-Plus Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Electroporation, Cell Culture, Quantitative RT-PCR, Expressing, Negative Control, Positive Control, Immunofluorescence, Staining

    Comparison of F9 expression, and hFIX antigen levels and activity between iPSC-parental-Heps and iPSC-insertion-Heps. a qRT-PCR showed F9 expression in undifferentiated iPSCs, iPSC-parental-Heps, and iPSC-insertion-Heps (n = 3 independent experiments for each sample). b Comparison of hFIX antigen levels in cell culture supernatant between iPSC-parental-Heps and iPSC-insertion-Heps. Student’s t test used. Data shown are mean ± SEM ( n = 8 in iPSC-parental-Heps group, n = 9 in iPSC-insertion-Heps group). c Comparison of hFIX activity in cell culture supernatant between iPSC-parental-Heps and iPSC-insertion-Heps. Student’s t test used. Data shown are mean ± SEM ( n = 4 in iPSC-parental-Heps group, n = 8 in iPSC-insertion-Heps group). d Two weeks after transplantation, hFIX antigen levels in mice plasma of the iPSC-insertion-Heps group, iPSC-parental-Heps group, and control group detected by ELISA. Student’s t test used. Data shown are mean ± SEM ( n = 5 in iPSC-insertion-Heps group and the control group, n = 4 in iPSC-parental-Heps group). e Two weeks after transplantation, human ALB was detected in mice #1, #2, #4, and #5 of the iPSC-insertion-Heps group by immunohistochemistry. All scale bars represent 50 μm. GAPDH glyceraldehyde 3-phosphate dehydrogenase, iPSC induced pluripotent cell, hFIX human factor IX, DAPI 4′,6-diamidino-2-phenylindole, ALB albumin

    Journal: Stem Cell Research & Therapy

    Article Title: Targeted genome engineering in human induced pluripotent stem cells from patients with hemophilia B using the CRISPR-Cas9 system

    doi: 10.1186/s13287-018-0839-8

    Figure Lengend Snippet: Comparison of F9 expression, and hFIX antigen levels and activity between iPSC-parental-Heps and iPSC-insertion-Heps. a qRT-PCR showed F9 expression in undifferentiated iPSCs, iPSC-parental-Heps, and iPSC-insertion-Heps (n = 3 independent experiments for each sample). b Comparison of hFIX antigen levels in cell culture supernatant between iPSC-parental-Heps and iPSC-insertion-Heps. Student’s t test used. Data shown are mean ± SEM ( n = 8 in iPSC-parental-Heps group, n = 9 in iPSC-insertion-Heps group). c Comparison of hFIX activity in cell culture supernatant between iPSC-parental-Heps and iPSC-insertion-Heps. Student’s t test used. Data shown are mean ± SEM ( n = 4 in iPSC-parental-Heps group, n = 8 in iPSC-insertion-Heps group). d Two weeks after transplantation, hFIX antigen levels in mice plasma of the iPSC-insertion-Heps group, iPSC-parental-Heps group, and control group detected by ELISA. Student’s t test used. Data shown are mean ± SEM ( n = 5 in iPSC-insertion-Heps group and the control group, n = 4 in iPSC-parental-Heps group). e Two weeks after transplantation, human ALB was detected in mice #1, #2, #4, and #5 of the iPSC-insertion-Heps group by immunohistochemistry. All scale bars represent 50 μm. GAPDH glyceraldehyde 3-phosphate dehydrogenase, iPSC induced pluripotent cell, hFIX human factor IX, DAPI 4′,6-diamidino-2-phenylindole, ALB albumin

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed in triplicate, using a Step-One-Plus Real-Time PCR system (Applied Biosystems, Foster City, CA, USA).

    Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Cell Culture, Transplantation Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

    Dose-Rate effect does not require cxcr4b in transplanted cells. A) WT zebrafish were irradiated with 20 Gy at either dose rate and transplanted with WKM from Tg( bactin :EGFP) WKM that was either heterozygous or homozygous for the odysseus ( cxcr4b ) mutation. 9 dpt, kidneys were harvested and analyzed by flow cytometry. Data shows pooled results from two biologically independent trials, n = 13–15 fish per group. Heterozygous: p = 0.0420, Homozygous mutant: p = 0.0141, (unpaired t-test). B) 1 day after radiation, samples enriched for renal tubules were collected from WT zebrafish and analyzed by qRT-PCR for sdf-1a expression. No significant differences were seen between dose-rate groups. Pooled results from two biological experiments, each with four fish per group, are shown. RQ: Relative Quantitation.

    Journal: PLoS ONE

    Article Title: Effect of Radiation Dose-Rate on Hematopoietic Cell Engraftment in Adult Zebrafish

    doi: 10.1371/journal.pone.0073745

    Figure Lengend Snippet: Dose-Rate effect does not require cxcr4b in transplanted cells. A) WT zebrafish were irradiated with 20 Gy at either dose rate and transplanted with WKM from Tg( bactin :EGFP) WKM that was either heterozygous or homozygous for the odysseus ( cxcr4b ) mutation. 9 dpt, kidneys were harvested and analyzed by flow cytometry. Data shows pooled results from two biologically independent trials, n = 13–15 fish per group. Heterozygous: p = 0.0420, Homozygous mutant: p = 0.0141, (unpaired t-test). B) 1 day after radiation, samples enriched for renal tubules were collected from WT zebrafish and analyzed by qRT-PCR for sdf-1a expression. No significant differences were seen between dose-rate groups. Pooled results from two biological experiments, each with four fish per group, are shown. RQ: Relative Quantitation.

    Article Snippet: RNA isolation and DNA removal were achieved with the RNeasy Mini Kit (Qiagen). cDNA was synthesized using SuperScript III 1st strand synthesis kit (Invitrogen). qRT-PCR was performed using Taqman Primers (listed in supplemental data) and reagents (Applied Biosystems). qRT-PCR reactions were performed in technical triplicates on a StepOnePlus qRT-PCR system (Applied Biosystems).

    Techniques: Irradiation, Mutagenesis, Flow Cytometry, Cytometry, Fluorescence In Situ Hybridization, Quantitative RT-PCR, Expressing, Quantitation Assay

    qRT-PCR of hematopoietic and non-hematopoietic kidney after radiation. 3/min (HDR) or a lower dose rate of 25 cGy/min (LDR), WT zebrafish kidneys were pooled and filtered to obtain samples enriched for either hematopoietic whole kidney marrow (WKM) (A), or renal tubules (B). 0 Gy control samples were harvested concurrently at each time point and set as biological references. Each biological trial was composed of kidneys from four fish per time point, and data summarizes pooled results from 2–3 independent biological trials per time point, per condition. Although LDR hematopoietic cells showed significantly higher p53 expression 2 dpr than HDR hematopoietic cells (.001

    Journal: PLoS ONE

    Article Title: Effect of Radiation Dose-Rate on Hematopoietic Cell Engraftment in Adult Zebrafish

    doi: 10.1371/journal.pone.0073745

    Figure Lengend Snippet: qRT-PCR of hematopoietic and non-hematopoietic kidney after radiation. 3/min (HDR) or a lower dose rate of 25 cGy/min (LDR), WT zebrafish kidneys were pooled and filtered to obtain samples enriched for either hematopoietic whole kidney marrow (WKM) (A), or renal tubules (B). 0 Gy control samples were harvested concurrently at each time point and set as biological references. Each biological trial was composed of kidneys from four fish per time point, and data summarizes pooled results from 2–3 independent biological trials per time point, per condition. Although LDR hematopoietic cells showed significantly higher p53 expression 2 dpr than HDR hematopoietic cells (.001

    Article Snippet: RNA isolation and DNA removal were achieved with the RNeasy Mini Kit (Qiagen). cDNA was synthesized using SuperScript III 1st strand synthesis kit (Invitrogen). qRT-PCR was performed using Taqman Primers (listed in supplemental data) and reagents (Applied Biosystems). qRT-PCR reactions were performed in technical triplicates on a StepOnePlus qRT-PCR system (Applied Biosystems).

    Techniques: Quantitative RT-PCR, Fluorescence In Situ Hybridization, Expressing

    Myelosuppression after 20Gy at 25 cGy/min vs 800cGy/min in non-transplanted fish. A, Flow cytometry analysis of hematopoietic cell recovery in the kidney on days 2, 6, and 20–21 following radiation at either 25cGy/min (LDR) or 800cGy/min (HDR), showing initial myelosuppression followed by hematopoietic recovery. Plot shows the means and SDs of pooled data from two independent biological trials, for a total of at least 7–10 fish per data point. 100,000 events were collected per sample. No significant differences were noted between dose-rate groups. B, qRT-PCR of hematopoietic cells 2 days post radiation. Data shows pooled results from two independent biological trials; each trial composed of 4 fish per group. No statistically significant differences between dose-rate groups were noted. RQ: Relative Quantitation.

    Journal: PLoS ONE

    Article Title: Effect of Radiation Dose-Rate on Hematopoietic Cell Engraftment in Adult Zebrafish

    doi: 10.1371/journal.pone.0073745

    Figure Lengend Snippet: Myelosuppression after 20Gy at 25 cGy/min vs 800cGy/min in non-transplanted fish. A, Flow cytometry analysis of hematopoietic cell recovery in the kidney on days 2, 6, and 20–21 following radiation at either 25cGy/min (LDR) or 800cGy/min (HDR), showing initial myelosuppression followed by hematopoietic recovery. Plot shows the means and SDs of pooled data from two independent biological trials, for a total of at least 7–10 fish per data point. 100,000 events were collected per sample. No significant differences were noted between dose-rate groups. B, qRT-PCR of hematopoietic cells 2 days post radiation. Data shows pooled results from two independent biological trials; each trial composed of 4 fish per group. No statistically significant differences between dose-rate groups were noted. RQ: Relative Quantitation.

    Article Snippet: RNA isolation and DNA removal were achieved with the RNeasy Mini Kit (Qiagen). cDNA was synthesized using SuperScript III 1st strand synthesis kit (Invitrogen). qRT-PCR was performed using Taqman Primers (listed in supplemental data) and reagents (Applied Biosystems). qRT-PCR reactions were performed in technical triplicates on a StepOnePlus qRT-PCR system (Applied Biosystems).

    Techniques: Fluorescence In Situ Hybridization, Flow Cytometry, Cytometry, Quantitative RT-PCR, Quantitation Assay