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pca25 cas9 sgrna egfp vector  (Addgene inc)


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    Addgene inc pca25 cas9 sgrna egfp vector
    Pca25 Cas9 Sgrna Egfp Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 2902 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cas9+egfp+vector/pSpCas9(BB)-2A-Puro+(PX459)+V2%2E0+(Plasmid+%2362988)/pm41857580-199-15-17
    Average 96 stars, based on 2902 article reviews
    pca25 cas9 sgrna egfp vector - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Enhancing the efficacy of glycolytic blockade in cancer cells via RAD51 inhibition
    Article Snippet: .. These guides were cloned into the Cas9-EGFP vector, pX330 (Addgene, Cat. no. 66582). .. These vectors were nucleofected into MEC1 cells using Amaxa Cell Line Nucleofector Kit V using program X-001, and according to manufacturer’s instructions (Lonza, Cat. no. VACA-1003).

    Article Title: High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells.
    Article Snippet: .. The target sequence was ordered as two complementary oligos (IDT) and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). .. For the generation of HHEX and LMX1A knockout organoids, gRNAs were selected using the WTSI website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). gRNAs used in this story are presented in Table S1.

    Article Title: Integration of metabolomics, genomics, and immune phenotypes reveals the causal roles of metabolites in disease
    Article Snippet: .. To generate FADS2 knockout organoids, gRNAs were selected using the Atum website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before [ ]. gRNAs used in this study were: Human colon organoids were transfected using these gRNAs cloned into the Cas9-EGFP vector, utilizing electroporation following a previously established protocol [ ]. .. One week after transfection, cells were sorted for EGFP positivity using a FACS-ARIA (BD Biosciences).

    Article Title: High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells.
    Article Snippet: The target sequence was ordered as two complementary oligos (IDT) and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). .. For the generation of HHEX and LMX1A knockout organoids, gRNAs were selected using the WTSI website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). gRNAs used in this story are presented in Table S1. ..

    Sequencing:

    Article Title: High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells.
    Article Snippet: .. The target sequence was ordered as two complementary oligos (IDT) and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). .. For the generation of HHEX and LMX1A knockout organoids, gRNAs were selected using the WTSI website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). gRNAs used in this story are presented in Table S1.

    Plasmid Preparation:

    Article Title: High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells.
    Article Snippet: .. The target sequence was ordered as two complementary oligos (IDT) and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). .. For the generation of HHEX and LMX1A knockout organoids, gRNAs were selected using the WTSI website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). gRNAs used in this story are presented in Table S1.

    Article Title: Integration of metabolomics, genomics, and immune phenotypes reveals the causal roles of metabolites in disease
    Article Snippet: .. To generate FADS2 knockout organoids, gRNAs were selected using the Atum website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before [ ]. gRNAs used in this study were: Human colon organoids were transfected using these gRNAs cloned into the Cas9-EGFP vector, utilizing electroporation following a previously established protocol [ ]. .. One week after transfection, cells were sorted for EGFP positivity using a FACS-ARIA (BD Biosciences).

    Article Title: High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells.
    Article Snippet: The target sequence was ordered as two complementary oligos (IDT) and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). .. For the generation of HHEX and LMX1A knockout organoids, gRNAs were selected using the WTSI website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). gRNAs used in this story are presented in Table S1. ..

    Knock-Out:

    Article Title: Integration of metabolomics, genomics, and immune phenotypes reveals the causal roles of metabolites in disease
    Article Snippet: .. To generate FADS2 knockout organoids, gRNAs were selected using the Atum website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before [ ]. gRNAs used in this study were: Human colon organoids were transfected using these gRNAs cloned into the Cas9-EGFP vector, utilizing electroporation following a previously established protocol [ ]. .. One week after transfection, cells were sorted for EGFP positivity using a FACS-ARIA (BD Biosciences).

    Article Title: High-Resolution mRNA and Secretome Atlas of Human Enteroendocrine Cells.
    Article Snippet: The target sequence was ordered as two complementary oligos (IDT) and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). .. For the generation of HHEX and LMX1A knockout organoids, gRNAs were selected using the WTSI website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before (Ran et al., 2013). gRNAs used in this story are presented in Table S1. ..

    Transfection:

    Article Title: Integration of metabolomics, genomics, and immune phenotypes reveals the causal roles of metabolites in disease
    Article Snippet: .. To generate FADS2 knockout organoids, gRNAs were selected using the Atum website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before [ ]. gRNAs used in this study were: Human colon organoids were transfected using these gRNAs cloned into the Cas9-EGFP vector, utilizing electroporation following a previously established protocol [ ]. .. One week after transfection, cells were sorted for EGFP positivity using a FACS-ARIA (BD Biosciences).

    Electroporation:

    Article Title: Integration of metabolomics, genomics, and immune phenotypes reveals the causal roles of metabolites in disease
    Article Snippet: .. To generate FADS2 knockout organoids, gRNAs were selected using the Atum website and cloned in the Cas9-EGFP vector (addgene plasmid #48138) following the protocol described before [ ]. gRNAs used in this study were: Human colon organoids were transfected using these gRNAs cloned into the Cas9-EGFP vector, utilizing electroporation following a previously established protocol [ ]. .. One week after transfection, cells were sorted for EGFP positivity using a FACS-ARIA (BD Biosciences).



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    Image Search Results


    Repair of 8-oxoG•A using a MUTYH lesion-specific plasmid reporter. ( A ) Schematic diagram of the fluorescent reporter for 8-oxoG•A repair. ( B ) Representative flow cytometry plots and gating schemes to quantify 8-oxoG•A repair in live cells using matching L111P MUTYH mutant cell lines as an example. The plots show compensated red fluorescence intensity ( y -axis) versus compensated EGFP fluorescence intensity ( x -axis) for four representative samples, left to right: unedited HEK293T cells, HEK293T cells that were transfected with the L111P gRNA, but produced no editing at the target site (null clone), heterozygous L111P MUTYH clone 1, and homozygous L111P MUTYH clone 1. 8-oxoG•A repair is quantified by calculating the percent of EGFP+ cells divided by the transfected, or mCherry+, cells. Scatter gates were applied to remove nonviable cells and doublets as shown in . Quadrant boundaries for analysis were set by using unedited HEK293T cells that were transfected with mCherry only or EGFP only plasmids. The numbers in each quadrant represent the percentage of cells within that population. “+’’s in the quadrants indicate the median EGFP fluorescence intensity of EGFP-positive cells. ( C ) Percentage of 8-oxoG•A repair in living cells harboring various MUTYH mutants. Values calculated as described in (B). Bars represent the average of n = 3 biological replicates (circles show clone 1, triangles show clone 2, and squares show clone 3). Error bars represent the standard deviation of the three biological replicates. Data were analyzed with unpaired, one-tailed, parametric t -tests (in which the heterozygous or homozygous lines were compared to their wild-type counterparts), and P- values are marked as follows: ns = P ≥ 0.05, not significant: * P ≤ .05, ** P ≤ .01, and **** P ≤ .0001 are significant.

    Journal: Nucleic Acids Research

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants

    doi: 10.1093/nar/gkaf037

    Figure Lengend Snippet: Repair of 8-oxoG•A using a MUTYH lesion-specific plasmid reporter. ( A ) Schematic diagram of the fluorescent reporter for 8-oxoG•A repair. ( B ) Representative flow cytometry plots and gating schemes to quantify 8-oxoG•A repair in live cells using matching L111P MUTYH mutant cell lines as an example. The plots show compensated red fluorescence intensity ( y -axis) versus compensated EGFP fluorescence intensity ( x -axis) for four representative samples, left to right: unedited HEK293T cells, HEK293T cells that were transfected with the L111P gRNA, but produced no editing at the target site (null clone), heterozygous L111P MUTYH clone 1, and homozygous L111P MUTYH clone 1. 8-oxoG•A repair is quantified by calculating the percent of EGFP+ cells divided by the transfected, or mCherry+, cells. Scatter gates were applied to remove nonviable cells and doublets as shown in . Quadrant boundaries for analysis were set by using unedited HEK293T cells that were transfected with mCherry only or EGFP only plasmids. The numbers in each quadrant represent the percentage of cells within that population. “+’’s in the quadrants indicate the median EGFP fluorescence intensity of EGFP-positive cells. ( C ) Percentage of 8-oxoG•A repair in living cells harboring various MUTYH mutants. Values calculated as described in (B). Bars represent the average of n = 3 biological replicates (circles show clone 1, triangles show clone 2, and squares show clone 3). Error bars represent the standard deviation of the three biological replicates. Data were analyzed with unpaired, one-tailed, parametric t -tests (in which the heterozygous or homozygous lines were compared to their wild-type counterparts), and P- values are marked as follows: ns = P ≥ 0.05, not significant: * P ≤ .05, ** P ≤ .01, and **** P ≤ .0001 are significant.

    Article Snippet: Guide RNA (gRNA) plasmids were cloned by site-directed mutagenesis using a 5′ tail in the forward primer to replace the 20 nt spacer region (Basic Protocol 1 [ ]; Streptococcus pyogenes Cas9 gRNA vector Addgene plasmid #47511).

    Techniques: Plasmid Preparation, Flow Cytometry, Mutagenesis, Fluorescence, Transfection, Produced, Standard Deviation, One-tailed Test