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streptococcus pyogenes cas9 grna vector addgene plasmid  (Addgene inc)


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    Addgene inc streptococcus pyogenes cas9 grna vector addgene plasmid
    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 <t>gRNA,</t> 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.
    Streptococcus Pyogenes Cas9 Grna Vector Addgene Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/streptococcus+pyogenes+cas9+grna+vector+addgene+plasmid/pFYF1320+EGFP+Site%231+(Plasmid+%2347511)/pmc11952967-51-29-34
    Average 93 stars, based on 30 article reviews
    streptococcus pyogenes cas9 grna vector addgene plasmid - by Bioz Stars, 2026-09
    93/100 stars

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    1) Product Images from "Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants"

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

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkaf037

    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.
    Figure Legend 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.

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

    Related Articles

    Clone Assay:

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants
    Article Snippet: All polymerase chain reaction (PCR) reactions were performed with Phusion DNA Green High-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) or Phusion U (Thermo Fisher, #F556L) where appropriate. .. 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). ..

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants.
    Article Snippet: M G A D ( H o G D ow nloaded from https://academ ic.oup.com /nar/article/53/6/gkaf037/8099200 by guest on 07 April 2025 aterials and methods eneral methods and cloning ll primers in this study were ordered through Integrated NA Technologies (IDT). .. All polymerase chain reaction PCR) reactions were performed with Phusion DNA Green igh-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) r Phusion U (Thermo Fisher, #F556L) where appropriate. uide 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 [ 57 ]; Streptococcus pyogenes Cas9 gRNA vector Addgene plasmid #47511). ..

    Mutagenesis:

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants
    Article Snippet: All polymerase chain reaction (PCR) reactions were performed with Phusion DNA Green High-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) or Phusion U (Thermo Fisher, #F556L) where appropriate. .. 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). ..

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants.
    Article Snippet: M G A D ( H o G D ow nloaded from https://academ ic.oup.com /nar/article/53/6/gkaf037/8099200 by guest on 07 April 2025 aterials and methods eneral methods and cloning ll primers in this study were ordered through Integrated NA Technologies (IDT). .. All polymerase chain reaction PCR) reactions were performed with Phusion DNA Green igh-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) r Phusion U (Thermo Fisher, #F556L) where appropriate. uide 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 [ 57 ]; Streptococcus pyogenes Cas9 gRNA vector Addgene plasmid #47511). ..

    Plasmid Preparation:

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants
    Article Snippet: All polymerase chain reaction (PCR) reactions were performed with Phusion DNA Green High-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) or Phusion U (Thermo Fisher, #F556L) where appropriate. .. 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). ..

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants.
    Article Snippet: M G A D ( H o G D ow nloaded from https://academ ic.oup.com /nar/article/53/6/gkaf037/8099200 by guest on 07 April 2025 aterials and methods eneral methods and cloning ll primers in this study were ordered through Integrated NA Technologies (IDT). .. All polymerase chain reaction PCR) reactions were performed with Phusion DNA Green igh-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) r Phusion U (Thermo Fisher, #F556L) where appropriate. uide 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 [ 57 ]; Streptococcus pyogenes Cas9 gRNA vector Addgene plasmid #47511). ..

    Polymerase Chain Reaction:

    Article Title: Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants.
    Article Snippet: M G A D ( H o G D ow nloaded from https://academ ic.oup.com /nar/article/53/6/gkaf037/8099200 by guest on 07 April 2025 aterials and methods eneral methods and cloning ll primers in this study were ordered through Integrated NA Technologies (IDT). .. All polymerase chain reaction PCR) reactions were performed with Phusion DNA Green igh-Fidelity Polymerase (Thermo Fisher Scientific, #F534L) r Phusion U (Thermo Fisher, #F556L) where appropriate. uide 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 [ 57 ]; Streptococcus pyogenes Cas9 gRNA vector Addgene plasmid #47511). ..



    Similar Products

    streptococcus pyogenes cas9 grna vector addgene plasmid  (Addgene inc)
    93
    Addgene inc streptococcus pyogenes cas9 grna vector addgene plasmid
    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 <t>gRNA,</t> 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.
    Streptococcus Pyogenes Cas9 Grna Vector Addgene Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/streptococcus+pyogenes+cas9+grna+vector+addgene+plasmid/pFYF1320+EGFP+Site%231+(Plasmid+%2347511)/pmc11952967-51-29-34
    Average 93 stars, based on 1 article reviews
    streptococcus pyogenes cas9 grna vector addgene plasmid - by Bioz Stars, 2026-09
    93/100 stars
    		  Buy from Supplier

    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