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Addgene inc oct4 tdtomato donor plasmid
Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of <t>OCT4-2A-tdTomato.</t> b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P
Oct4 Tdtomato Donor Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/oct4 tdtomato donor plasmid/product/Addgene inc
Average 92 stars, based on 19 article reviews
Price from $9.99 to $1999.99
oct4 tdtomato donor plasmid - by Bioz Stars, 2020-08
92/100 stars

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1) Product Images from "Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells"

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

Journal: Nature Communications

doi: 10.1038/s41467-018-03760-5

Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P
Figure Legend Snippet: Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

Techniques Used: CRISPR, Knock-In, Selection, FACS, Transfection, Plasmid Preparation, Two Tailed Test

2) Product Images from "Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells"

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

Journal: Nature Communications

doi: 10.1038/s41467-018-03760-5

Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P
Figure Legend Snippet: Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

Techniques Used: CRISPR, Knock-In, Selection, FACS, Transfection, Plasmid Preparation, Two Tailed Test

3) Product Images from "Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells"

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

Journal: Nature Communications

doi: 10.1038/s41467-018-03760-5

Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P
Figure Legend Snippet: Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

Techniques Used: CRISPR, Knock-In, Selection, FACS, Transfection, Plasmid Preparation, Two Tailed Test

4) Product Images from "Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells"

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

Journal: Nature Communications

doi: 10.1038/s41467-018-03760-5

Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P
Figure Legend Snippet: Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

Techniques Used: CRISPR, Knock-In, Selection, FACS, Transfection, Plasmid Preparation, Two Tailed Test

Related Articles

Transfection:

Article Title: Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang
Article Snippet: .. The mixture (300 μL) was added to 1 mL DMEM media in which 5 × 105 cells were plated 1 day before transfection, and cells were grown in the presence of the mixture for 48 h. After incubation, cells were harvested, and genomic DNA was prepared either manually using a PureHelixTM Genomic DNA Preparation Kit (NanoHelix) or using a MaxwellTM RSC nucleic acid isolation workstation (Promega). pSpCas9(BB)-2A-GFP (PX458), pY010(pcDNA3.1-hAsCpf1), and pY016 (pcDNA3.1-hLbCpf1) were gifts from Feng Zhang (Addgene plasmid #48138, #69982, #69988, respectively). ..

Amplification:

Article Title: Assessment of Cas12a‐mediated gene editing efficiency in plants
Article Snippet: .. Plasmids pY010 (pcDNA3.1‐hAsCpf1, Addgene plasmid # 69982) and pY016 (pcDNA3.1‐hLbCpf1, Addgene plasmid # 69988), kindly provided by Feng Zhang Laboratory, served as a template for the construction of GB1438 and GB1439 by PCR amplification, using the Phusion High‐Fidelity DNA Polymerase (Thermo Scientific). .. GB1442 and GB1443 were also obtained by PCR amplification of GB1001.

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells
Article Snippet: .. For OCT4-tdTomato donor plasmid, OCT4-2A-mOrange donor plasmid (Addgene, Plasmid #66986) was PCR amplified without mOrange sequence as the backbone, and the tdTomato sequence was amplified and linked with backbone by using Gibson Assembly kit (New England Biolabs). .. All vectors were checked by Sanger sequencing.

Polymerase Chain Reaction:

Article Title: Assessment of Cas12a‐mediated gene editing efficiency in plants
Article Snippet: .. Plasmids pY010 (pcDNA3.1‐hAsCpf1, Addgene plasmid # 69982) and pY016 (pcDNA3.1‐hLbCpf1, Addgene plasmid # 69988), kindly provided by Feng Zhang Laboratory, served as a template for the construction of GB1438 and GB1439 by PCR amplification, using the Phusion High‐Fidelity DNA Polymerase (Thermo Scientific). .. GB1442 and GB1443 were also obtained by PCR amplification of GB1001.

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells
Article Snippet: .. For OCT4-tdTomato donor plasmid, OCT4-2A-mOrange donor plasmid (Addgene, Plasmid #66986) was PCR amplified without mOrange sequence as the backbone, and the tdTomato sequence was amplified and linked with backbone by using Gibson Assembly kit (New England Biolabs). .. All vectors were checked by Sanger sequencing.

Isolation:

Article Title: Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang
Article Snippet: .. The mixture (300 μL) was added to 1 mL DMEM media in which 5 × 105 cells were plated 1 day before transfection, and cells were grown in the presence of the mixture for 48 h. After incubation, cells were harvested, and genomic DNA was prepared either manually using a PureHelixTM Genomic DNA Preparation Kit (NanoHelix) or using a MaxwellTM RSC nucleic acid isolation workstation (Promega). pSpCas9(BB)-2A-GFP (PX458), pY010(pcDNA3.1-hAsCpf1), and pY016 (pcDNA3.1-hLbCpf1) were gifts from Feng Zhang (Addgene plasmid #48138, #69982, #69988, respectively). ..

Knock-In:

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells
Article Snippet: .. For knockin experiments, plasmid mixture was prepared with 3 μg of pcDNA3.1-hLbCpf1, 3 μg of pCpfcr-crRNA, and 4 μg of OCT4-tdTomato donor or OCT4-2A-eGFP-PGK-Puro donor (Addgene, Plasmid #31938) or 120-nt ssODN template. .. For double knockin, plasmid mixture was prepared with 4 μg of pcDNA3.1-hLbCpf1, 3 μg of pCpfcr-OCT4-crRNA1, 3 μg of pCpfcr-ALBUMIN-crRNA1, 4 μg of OCT4-tdTomato donor, and 4 μg of ALBUMIN-2A-tdTomato donor. hPSCs were dissociated into single cells by using Accutase.

Incubation:

Article Title: Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang
Article Snippet: .. The mixture (300 μL) was added to 1 mL DMEM media in which 5 × 105 cells were plated 1 day before transfection, and cells were grown in the presence of the mixture for 48 h. After incubation, cells were harvested, and genomic DNA was prepared either manually using a PureHelixTM Genomic DNA Preparation Kit (NanoHelix) or using a MaxwellTM RSC nucleic acid isolation workstation (Promega). pSpCas9(BB)-2A-GFP (PX458), pY010(pcDNA3.1-hAsCpf1), and pY016 (pcDNA3.1-hLbCpf1) were gifts from Feng Zhang (Addgene plasmid #48138, #69982, #69988, respectively). ..

Sequencing:

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells
Article Snippet: .. For OCT4-tdTomato donor plasmid, OCT4-2A-mOrange donor plasmid (Addgene, Plasmid #66986) was PCR amplified without mOrange sequence as the backbone, and the tdTomato sequence was amplified and linked with backbone by using Gibson Assembly kit (New England Biolabs). .. All vectors were checked by Sanger sequencing.

Plasmid Preparation:

Article Title: Assessment of Cas12a‐mediated gene editing efficiency in plants
Article Snippet: .. Plasmids pY010 (pcDNA3.1‐hAsCpf1, Addgene plasmid # 69982) and pY016 (pcDNA3.1‐hLbCpf1, Addgene plasmid # 69988), kindly provided by Feng Zhang Laboratory, served as a template for the construction of GB1438 and GB1439 by PCR amplification, using the Phusion High‐Fidelity DNA Polymerase (Thermo Scientific). .. GB1442 and GB1443 were also obtained by PCR amplification of GB1001.

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells
Article Snippet: .. For knockin experiments, plasmid mixture was prepared with 3 μg of pcDNA3.1-hLbCpf1, 3 μg of pCpfcr-crRNA, and 4 μg of OCT4-tdTomato donor or OCT4-2A-eGFP-PGK-Puro donor (Addgene, Plasmid #31938) or 120-nt ssODN template. .. For double knockin, plasmid mixture was prepared with 4 μg of pcDNA3.1-hLbCpf1, 3 μg of pCpfcr-OCT4-crRNA1, 3 μg of pCpfcr-ALBUMIN-crRNA1, 4 μg of OCT4-tdTomato donor, and 4 μg of ALBUMIN-2A-tdTomato donor. hPSCs were dissociated into single cells by using Accutase.

Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells
Article Snippet: .. For OCT4-tdTomato donor plasmid, OCT4-2A-mOrange donor plasmid (Addgene, Plasmid #66986) was PCR amplified without mOrange sequence as the backbone, and the tdTomato sequence was amplified and linked with backbone by using Gibson Assembly kit (New England Biolabs). .. All vectors were checked by Sanger sequencing.

Article Title: Cpf1 proteins excise CRISPR RNAs from mRNA transcripts in mammalian cells
Article Snippet: .. Plasmids Wild-type LbCpf1 plasmid, pY016 (pcDNA3.1-hLbCpf1, Addgene plasmid # 69988), and AsCpf1 plasmid, pY010 (pcDNA3.1-hAsCpf1, Addgene plasmid # 69982) were gifts from the laboratory of Dr. Feng Zhang . .. LbCpf1 mutations (H759A, K768A, K785A, HKK/AAA, F789A, D832A, E925A) were introduced by synthesis of the LbCpf1 fragment on pY016 plasmid between Asc I and Ale I restriction sites.

Article Title: Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang
Article Snippet: .. The mixture (300 μL) was added to 1 mL DMEM media in which 5 × 105 cells were plated 1 day before transfection, and cells were grown in the presence of the mixture for 48 h. After incubation, cells were harvested, and genomic DNA was prepared either manually using a PureHelixTM Genomic DNA Preparation Kit (NanoHelix) or using a MaxwellTM RSC nucleic acid isolation workstation (Promega). pSpCas9(BB)-2A-GFP (PX458), pY010(pcDNA3.1-hAsCpf1), and pY016 (pcDNA3.1-hLbCpf1) were gifts from Feng Zhang (Addgene plasmid #48138, #69982, #69988, respectively). ..

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    Addgene inc oct4 tdtomato donor plasmid
    Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of <t>OCT4-2A-tdTomato.</t> b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P
    Oct4 Tdtomato Donor Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oct4 tdtomato donor plasmid/product/Addgene inc
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    oct4 tdtomato donor plasmid - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

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    Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

    Journal: Nature Communications

    Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

    doi: 10.1038/s41467-018-03760-5

    Figure Lengend Snippet: Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

    Article Snippet: For OCT4-tdTomato donor plasmid, OCT4-2A-mOrange donor plasmid (Addgene, Plasmid #66986) was PCR amplified without mOrange sequence as the backbone, and the tdTomato sequence was amplified and linked with backbone by using Gibson Assembly kit (New England Biolabs).

    Techniques: CRISPR, Knock-In, Selection, FACS, Transfection, Plasmid Preparation, Two Tailed Test

    Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

    Journal: Nature Communications

    Article Title: Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

    doi: 10.1038/s41467-018-03760-5

    Figure Lengend Snippet: Small molecules significantly promote CRISPR-Cpf1-mediated generation of knockin hPSC lines without drug selection. a A scheme of the targeting strategy of OCT4-2A-tdTomato. b FACS analysis for OCT4-tdTomato-positive cells after transfection of the OCT4-tdTomato donor plasmid and the OCT4-targeting crRNA into hPSCs. A representative FACS result of samples treated with DMSO, VE-822, and AZD-7762 was shown. c Addition of VE-822 and AZD-7762 increased the percentage of OCT4-tdTomato-positive cells. n = 3 experiments. Statistical significance calculated using two-tailed Student’s t -test, compared to DMSO controls. * P

    Article Snippet: For OCT4-tdTomato donor plasmid, OCT4-2A-mOrange donor plasmid (Addgene, Plasmid #66986) was PCR amplified without mOrange sequence as the backbone, and the tdTomato sequence was amplified and linked with backbone by using Gibson Assembly kit (New England Biolabs).

    Techniques: CRISPR, Knock-In, Selection, FACS, Transfection, Plasmid Preparation, Two Tailed Test