sgrna guided cas9  (New England Biolabs)


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    Name:
    EnGen Spy Cas9 NLS
    Description:
    EnGen Spy Cas9 NLS 2 000 pmol
    Catalog Number:
    M0646M
    Price:
    600
    Category:
    Other Endonucleases
    Size:
    2 000 pmol
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    Structured Review

    New England Biolabs sgrna guided cas9
    EnGen Spy Cas9 NLS
    EnGen Spy Cas9 NLS 2 000 pmol
    https://www.bioz.com/result/sgrna guided cas9/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sgrna guided cas9 - by Bioz Stars, 2021-06
    99/100 stars

    Images

    1) Product Images from "The Sigma-2 Receptor and Progesterone Receptor Membrane Component 1 are Different Binding Sites Derived From Independent Genes"

    Article Title: The Sigma-2 Receptor and Progesterone Receptor Membrane Component 1 are Different Binding Sites Derived From Independent Genes

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2015.10.017

    PGRMC1 knockout using a CRISPR/Cas9 approach. A. Schematic of the PGRMC1 sgRNA/Cas9-expressing lentiviral constructs for knocking out PGRMC1. Gray areas show the three candidate sgRNA sequences in the exon-1 of the PGRMC1 gene, of which two were used for the CRISPR/Cas9 constructs. B. A representative DNA gel of control and PGRMC1 knockout (clones 38 and 207) NSC34 cells verifying the Cas9 cleavage of the genomic DNA. Control refers to the NSC34 cells transfected with the control vector expressing Cas9 but not an sgRNA. C. Western blotting detection of PGRMC1 in control and PGRMC1 knockout (clones 38 and 207) NSC34 cells.
    Figure Legend Snippet: PGRMC1 knockout using a CRISPR/Cas9 approach. A. Schematic of the PGRMC1 sgRNA/Cas9-expressing lentiviral constructs for knocking out PGRMC1. Gray areas show the three candidate sgRNA sequences in the exon-1 of the PGRMC1 gene, of which two were used for the CRISPR/Cas9 constructs. B. A representative DNA gel of control and PGRMC1 knockout (clones 38 and 207) NSC34 cells verifying the Cas9 cleavage of the genomic DNA. Control refers to the NSC34 cells transfected with the control vector expressing Cas9 but not an sgRNA. C. Western blotting detection of PGRMC1 in control and PGRMC1 knockout (clones 38 and 207) NSC34 cells.

    Techniques Used: Knock-Out, CRISPR, Expressing, Construct, Transfection, Plasmid Preparation, Western Blot

    Eliminating PGRMC1 protein does not alter [ 3 H]-DTG binding to the S2R in cell membranes. A. A representative of [ 3 H]-DTG saturation binding in membranes prepared from control and PGRMC1 knockout (clones 38 and 207) NSC34 cells, (+)-pentazocine (100 nM) was included to mask [ 3 H]-DTG binding to the S1R such that [ 3 H]-DTG would be bound only to the S2R and measured as specific S2R binding. Nonspecific binding was measured (by adding haloperidol) and subtracted. Control refers to the NSC34 cells transfected with the control vector for the expression of Cas9 but not an sgRNA. B. Statistics. Maximum binding (B max ) and equilibrium dissociation constants (K D ) for [ 3 H]-DTG were calculated using a Prizm software and reported as mean ± SEM of three separate experiments each performed in triplicates. Not significant (n.s.).
    Figure Legend Snippet: Eliminating PGRMC1 protein does not alter [ 3 H]-DTG binding to the S2R in cell membranes. A. A representative of [ 3 H]-DTG saturation binding in membranes prepared from control and PGRMC1 knockout (clones 38 and 207) NSC34 cells, (+)-pentazocine (100 nM) was included to mask [ 3 H]-DTG binding to the S1R such that [ 3 H]-DTG would be bound only to the S2R and measured as specific S2R binding. Nonspecific binding was measured (by adding haloperidol) and subtracted. Control refers to the NSC34 cells transfected with the control vector for the expression of Cas9 but not an sgRNA. B. Statistics. Maximum binding (B max ) and equilibrium dissociation constants (K D ) for [ 3 H]-DTG were calculated using a Prizm software and reported as mean ± SEM of three separate experiments each performed in triplicates. Not significant (n.s.).

    Techniques Used: Binding Assay, Knock-Out, Transfection, Plasmid Preparation, Expressing, Software

    Related Articles

    Knock-Out:

    Article Title: Toxoplasma gondii co-opts the unfolded protein response to enhance migration and dissemination of infected host cells
    Article Snippet: Cultures of DCs and J774.1 macrophages were cultivated in Roswell Park Memorial Institute (RPMI) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco/Invitrogen) and penicillin/streptomycin at 37°C with 5% CO2 and infected as described above. .. Generation of IRE1 knockout cells Disruption of the IRE1 gene in MEF cells was carried out using the CRISPR/Cas9 method [ ]. ..

    CRISPR:

    Article Title: Toxoplasma gondii co-opts the unfolded protein response to enhance migration and dissemination of infected host cells
    Article Snippet: Cultures of DCs and J774.1 macrophages were cultivated in Roswell Park Memorial Institute (RPMI) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco/Invitrogen) and penicillin/streptomycin at 37°C with 5% CO2 and infected as described above. .. Generation of IRE1 knockout cells Disruption of the IRE1 gene in MEF cells was carried out using the CRISPR/Cas9 method [ ]. ..

    Article Title: A fast Myh super enhancer dictates adult muscle fiber phenotype through competitive interactions with the fast Myh genes
    Article Snippet: .. Mouse generation by CRISPR/Cas9 SgRNA and Cas9 purified protein were produced by the TACGENE platform. ..

    Article Title: The use of CRISPR-Cas Selective Amplicon Sequencing (CCSAS) to reveal the eukaryotic microbiome of metazoans
    Article Snippet: The reaction was incubated at 37°C for 4 h in a thermocycler, and then followed by 70°C for 10 min to deactivate the CRISPR-Cas9. .. The reaction was incubated at 37°C for 4 h in a thermocycler, and then followed by 70°C for 10 min to deactivate the CRISPR-Cas9. .. For each sample, in parallel with the CRISPR-Cas9 treatment we also prepared the reaction without CRISPR-Cas9 treatment, in which Cas9 nuclease and sgRNA were replaced with molecular grade ultrapure water (Invitrogen).

    Article Title: Genome-wide CRISPR screens of oral squamous cell carcinoma reveal fitness genes in the Hippo pathway
    Article Snippet: .. Validation sgRNA design and cloning To validate the results obtained from the screen, individual targeted genetic knockouts were generated using CRISPR/Cas9 and plasmid expressing sgRNAs targeting the gene of interest. .. Two sgRNA sequences were used for each target gene, one was selected from the CRISPR library v1 while another sequence was designed using the Genetic Perturbation Platform (GPP) web portal ( https://portals.broadinstitute.org/gpp/public/analysis-tools/sgrna-design ).

    Article Title: Rotavirus induces intercellular calcium waves through ADP signaling
    Article Snippet: MA104-GCaMP6s cells (pLVX-IRES-Hygro) were transduced with the P2Y1 CRISPR/Cas9-expressing construct and at 72 hrs post-transduction the cells were passaged in the presence of 50 μg/mL hygromycin B and 10 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. The J2-derived jHIE-GCaMP6s(hygro) enteroid cultures described above were transduced with the CRISPR/Cas9-expressing constructs to make two separate enteroid lines and grown with 50 μg/mL hygromycin B and 1 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. Genotyping was performed on whole genomic DNA prepared using DirectPCR (Viagen) with Proteinase K (Qiagen) according to manufacturer’s instructions.

    Article Title: Genetic variation among wMel strains of Wolbachia pipientis differentially rescues a bag of marbles partial loss of function mutant in Drosophila melanogaster
    Article Snippet: We backcrossed females of all CRISPR/Cas9 mutants to w1118 isogenic males for three generations, and then crossed the mutants to the w1118 ; TM2/TM6 line to maintain the bam mutants over the TM6 balancer. .. All CRISPR/Cas9 edits in the lines were confirmed by Sanger sequencing (Cornell BRC Genomics Facility). .. PCR assays to detect Wolbachia To test for the presence of Wolbachia , we used the Zymo quick DNA miniprep kit to prepare DNA from three replicate samples each with three female flies.

    Article Title: Rotavirus induces intercellular calcium waves through ADP signaling
    Article Snippet: .. MA104-GCaMP6s cells (pLVX-IRES-Hygro) were transduced with the P2Y1 CRISPR/Cas9-expressing construct and at 72 hrs post-transduction the cells were passaged in the presence of 50 μg/mL hygromycin B and 10 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. The J2-derived jHIE-GCaMP6s(hygro) enteroid cultures described above were transduced with the CRISPR/Cas9-expressing constructs to make two separate enteroid lines and grown with 50 μg/mL hygromycin B and 1 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9.

    Purification:

    Article Title: A fast Myh super enhancer dictates adult muscle fiber phenotype through competitive interactions with the fast Myh genes
    Article Snippet: .. Mouse generation by CRISPR/Cas9 SgRNA and Cas9 purified protein were produced by the TACGENE platform. ..

    Produced:

    Article Title: A fast Myh super enhancer dictates adult muscle fiber phenotype through competitive interactions with the fast Myh genes
    Article Snippet: .. Mouse generation by CRISPR/Cas9 SgRNA and Cas9 purified protein were produced by the TACGENE platform. ..

    Incubation:

    Article Title: The use of CRISPR-Cas Selective Amplicon Sequencing (CCSAS) to reveal the eukaryotic microbiome of metazoans
    Article Snippet: The reaction was incubated at 37°C for 4 h in a thermocycler, and then followed by 70°C for 10 min to deactivate the CRISPR-Cas9. .. The reaction was incubated at 37°C for 4 h in a thermocycler, and then followed by 70°C for 10 min to deactivate the CRISPR-Cas9. .. For each sample, in parallel with the CRISPR-Cas9 treatment we also prepared the reaction without CRISPR-Cas9 treatment, in which Cas9 nuclease and sgRNA were replaced with molecular grade ultrapure water (Invitrogen).

    Clone Assay:

    Article Title: Genome-wide CRISPR screens of oral squamous cell carcinoma reveal fitness genes in the Hippo pathway
    Article Snippet: .. Validation sgRNA design and cloning To validate the results obtained from the screen, individual targeted genetic knockouts were generated using CRISPR/Cas9 and plasmid expressing sgRNAs targeting the gene of interest. .. Two sgRNA sequences were used for each target gene, one was selected from the CRISPR library v1 while another sequence was designed using the Genetic Perturbation Platform (GPP) web portal ( https://portals.broadinstitute.org/gpp/public/analysis-tools/sgrna-design ).

    Generated:

    Article Title: Genome-wide CRISPR screens of oral squamous cell carcinoma reveal fitness genes in the Hippo pathway
    Article Snippet: .. Validation sgRNA design and cloning To validate the results obtained from the screen, individual targeted genetic knockouts were generated using CRISPR/Cas9 and plasmid expressing sgRNAs targeting the gene of interest. .. Two sgRNA sequences were used for each target gene, one was selected from the CRISPR library v1 while another sequence was designed using the Genetic Perturbation Platform (GPP) web portal ( https://portals.broadinstitute.org/gpp/public/analysis-tools/sgrna-design ).

    Plasmid Preparation:

    Article Title: Genome-wide CRISPR screens of oral squamous cell carcinoma reveal fitness genes in the Hippo pathway
    Article Snippet: .. Validation sgRNA design and cloning To validate the results obtained from the screen, individual targeted genetic knockouts were generated using CRISPR/Cas9 and plasmid expressing sgRNAs targeting the gene of interest. .. Two sgRNA sequences were used for each target gene, one was selected from the CRISPR library v1 while another sequence was designed using the Genetic Perturbation Platform (GPP) web portal ( https://portals.broadinstitute.org/gpp/public/analysis-tools/sgrna-design ).

    Expressing:

    Article Title: Genome-wide CRISPR screens of oral squamous cell carcinoma reveal fitness genes in the Hippo pathway
    Article Snippet: .. Validation sgRNA design and cloning To validate the results obtained from the screen, individual targeted genetic knockouts were generated using CRISPR/Cas9 and plasmid expressing sgRNAs targeting the gene of interest. .. Two sgRNA sequences were used for each target gene, one was selected from the CRISPR library v1 while another sequence was designed using the Genetic Perturbation Platform (GPP) web portal ( https://portals.broadinstitute.org/gpp/public/analysis-tools/sgrna-design ).

    Transduction:

    Article Title: Rotavirus induces intercellular calcium waves through ADP signaling
    Article Snippet: MA104-GCaMP6s cells (pLVX-IRES-Hygro) were transduced with the P2Y1 CRISPR/Cas9-expressing construct and at 72 hrs post-transduction the cells were passaged in the presence of 50 μg/mL hygromycin B and 10 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. The J2-derived jHIE-GCaMP6s(hygro) enteroid cultures described above were transduced with the CRISPR/Cas9-expressing constructs to make two separate enteroid lines and grown with 50 μg/mL hygromycin B and 1 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. Genotyping was performed on whole genomic DNA prepared using DirectPCR (Viagen) with Proteinase K (Qiagen) according to manufacturer’s instructions.

    Article Title: Rotavirus induces intercellular calcium waves through ADP signaling
    Article Snippet: .. MA104-GCaMP6s cells (pLVX-IRES-Hygro) were transduced with the P2Y1 CRISPR/Cas9-expressing construct and at 72 hrs post-transduction the cells were passaged in the presence of 50 μg/mL hygromycin B and 10 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. The J2-derived jHIE-GCaMP6s(hygro) enteroid cultures described above were transduced with the CRISPR/Cas9-expressing constructs to make two separate enteroid lines and grown with 50 μg/mL hygromycin B and 1 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9.

    Construct:

    Article Title: Rotavirus induces intercellular calcium waves through ADP signaling
    Article Snippet: MA104-GCaMP6s cells (pLVX-IRES-Hygro) were transduced with the P2Y1 CRISPR/Cas9-expressing construct and at 72 hrs post-transduction the cells were passaged in the presence of 50 μg/mL hygromycin B and 10 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. The J2-derived jHIE-GCaMP6s(hygro) enteroid cultures described above were transduced with the CRISPR/Cas9-expressing constructs to make two separate enteroid lines and grown with 50 μg/mL hygromycin B and 1 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. Genotyping was performed on whole genomic DNA prepared using DirectPCR (Viagen) with Proteinase K (Qiagen) according to manufacturer’s instructions.

    Article Title: Rotavirus induces intercellular calcium waves through ADP signaling
    Article Snippet: .. MA104-GCaMP6s cells (pLVX-IRES-Hygro) were transduced with the P2Y1 CRISPR/Cas9-expressing construct and at 72 hrs post-transduction the cells were passaged in the presence of 50 μg/mL hygromycin B and 10 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9. .. The J2-derived jHIE-GCaMP6s(hygro) enteroid cultures described above were transduced with the CRISPR/Cas9-expressing constructs to make two separate enteroid lines and grown with 50 μg/mL hygromycin B and 1 μg/mL puromycin to select for co-expression of GCaMP6s and CRISPR/Cas9.

    Sequencing:

    Article Title: Genetic variation among wMel strains of Wolbachia pipientis differentially rescues a bag of marbles partial loss of function mutant in Drosophila melanogaster
    Article Snippet: We backcrossed females of all CRISPR/Cas9 mutants to w1118 isogenic males for three generations, and then crossed the mutants to the w1118 ; TM2/TM6 line to maintain the bam mutants over the TM6 balancer. .. All CRISPR/Cas9 edits in the lines were confirmed by Sanger sequencing (Cornell BRC Genomics Facility). .. PCR assays to detect Wolbachia To test for the presence of Wolbachia , we used the Zymo quick DNA miniprep kit to prepare DNA from three replicate samples each with three female flies.

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    New England Biolabs sgrna guided cas9
    PGRMC1 knockout using a <t>CRISPR/Cas9</t> approach. A. Schematic of the PGRMC1 <t>sgRNA/Cas9-expressing</t> lentiviral constructs for knocking out PGRMC1. Gray areas show the three candidate sgRNA sequences in the exon-1 of the PGRMC1 gene, of which two were used for the CRISPR/Cas9 constructs. B. A representative DNA gel of control and PGRMC1 knockout (clones 38 and 207) NSC34 cells verifying the Cas9 cleavage of the genomic DNA. Control refers to the NSC34 cells transfected with the control vector expressing Cas9 but not an sgRNA. C. Western blotting detection of PGRMC1 in control and PGRMC1 knockout (clones 38 and 207) NSC34 cells.
    Sgrna Guided Cas9, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sgrna guided cas9/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sgrna guided cas9 - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

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    PGRMC1 knockout using a CRISPR/Cas9 approach. A. Schematic of the PGRMC1 sgRNA/Cas9-expressing lentiviral constructs for knocking out PGRMC1. Gray areas show the three candidate sgRNA sequences in the exon-1 of the PGRMC1 gene, of which two were used for the CRISPR/Cas9 constructs. B. A representative DNA gel of control and PGRMC1 knockout (clones 38 and 207) NSC34 cells verifying the Cas9 cleavage of the genomic DNA. Control refers to the NSC34 cells transfected with the control vector expressing Cas9 but not an sgRNA. C. Western blotting detection of PGRMC1 in control and PGRMC1 knockout (clones 38 and 207) NSC34 cells.

    Journal: EBioMedicine

    Article Title: The Sigma-2 Receptor and Progesterone Receptor Membrane Component 1 are Different Binding Sites Derived From Independent Genes

    doi: 10.1016/j.ebiom.2015.10.017

    Figure Lengend Snippet: PGRMC1 knockout using a CRISPR/Cas9 approach. A. Schematic of the PGRMC1 sgRNA/Cas9-expressing lentiviral constructs for knocking out PGRMC1. Gray areas show the three candidate sgRNA sequences in the exon-1 of the PGRMC1 gene, of which two were used for the CRISPR/Cas9 constructs. B. A representative DNA gel of control and PGRMC1 knockout (clones 38 and 207) NSC34 cells verifying the Cas9 cleavage of the genomic DNA. Control refers to the NSC34 cells transfected with the control vector expressing Cas9 but not an sgRNA. C. Western blotting detection of PGRMC1 in control and PGRMC1 knockout (clones 38 and 207) NSC34 cells.

    Article Snippet: In order to assess the efficiency of sgRNA-guided Cas9 cutting in the PGRMC1 genomic sequence (exon-1), genomic DNA was extracted for PCR amplification of the specific region including the sgRNA/Cas9 excision site.

    Techniques: Knock-Out, CRISPR, Expressing, Construct, Transfection, Plasmid Preparation, Western Blot

    Eliminating PGRMC1 protein does not alter [ 3 H]-DTG binding to the S2R in cell membranes. A. A representative of [ 3 H]-DTG saturation binding in membranes prepared from control and PGRMC1 knockout (clones 38 and 207) NSC34 cells, (+)-pentazocine (100 nM) was included to mask [ 3 H]-DTG binding to the S1R such that [ 3 H]-DTG would be bound only to the S2R and measured as specific S2R binding. Nonspecific binding was measured (by adding haloperidol) and subtracted. Control refers to the NSC34 cells transfected with the control vector for the expression of Cas9 but not an sgRNA. B. Statistics. Maximum binding (B max ) and equilibrium dissociation constants (K D ) for [ 3 H]-DTG were calculated using a Prizm software and reported as mean ± SEM of three separate experiments each performed in triplicates. Not significant (n.s.).

    Journal: EBioMedicine

    Article Title: The Sigma-2 Receptor and Progesterone Receptor Membrane Component 1 are Different Binding Sites Derived From Independent Genes

    doi: 10.1016/j.ebiom.2015.10.017

    Figure Lengend Snippet: Eliminating PGRMC1 protein does not alter [ 3 H]-DTG binding to the S2R in cell membranes. A. A representative of [ 3 H]-DTG saturation binding in membranes prepared from control and PGRMC1 knockout (clones 38 and 207) NSC34 cells, (+)-pentazocine (100 nM) was included to mask [ 3 H]-DTG binding to the S1R such that [ 3 H]-DTG would be bound only to the S2R and measured as specific S2R binding. Nonspecific binding was measured (by adding haloperidol) and subtracted. Control refers to the NSC34 cells transfected with the control vector for the expression of Cas9 but not an sgRNA. B. Statistics. Maximum binding (B max ) and equilibrium dissociation constants (K D ) for [ 3 H]-DTG were calculated using a Prizm software and reported as mean ± SEM of three separate experiments each performed in triplicates. Not significant (n.s.).

    Article Snippet: In order to assess the efficiency of sgRNA-guided Cas9 cutting in the PGRMC1 genomic sequence (exon-1), genomic DNA was extracted for PCR amplification of the specific region including the sgRNA/Cas9 excision site.

    Techniques: Binding Assay, Knock-Out, Transfection, Plasmid Preparation, Expressing, Software