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pshs306  (Addgene inc)


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    Structured Review

    Addgene inc pshs306
    Pshs306, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+pshs306/pSHS306+-+Bacterial+expression+plasmid+for+SpCas9%2C+HNH+FRET+variant+(Plasmid+%2387350)/pm34767923-71-7-9
    Average 90 stars, based on 2 article reviews
    pshs306 - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Engineered CRISPR/Cas9 enzymes improve discrimination by slowing DNA cleavage to allow release of off-target DNA
    Article Snippet: .. Plasmid pMJ806 containing Streptococcus pyogenes Cas9 and plasmid pSHS306 containing the HNH FRET variant of Streptococcus pyogenes Cas9 were obtained from Addgene (Cambridge, MA). ..

    Variant Assay:

    Article Title: Engineered CRISPR/Cas9 enzymes improve discrimination by slowing DNA cleavage to allow release of off-target DNA
    Article Snippet: .. Plasmid pMJ806 containing Streptococcus pyogenes Cas9 and plasmid pSHS306 containing the HNH FRET variant of Streptococcus pyogenes Cas9 were obtained from Addgene (Cambridge, MA). ..



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    R-loop formation decay rates were measured by the monitoring the fluorescence increase from tC o (position −1 in the non-target strand) as a function of time after mixing enzyme (28 nM) with DNA (10 nM) in the presence of 10 mM Mg 2+ . Data were fit to a double exponential function to get the decay rates shown. a, b SpCas9 (28 nm) with on-target ( a ) and off-target ( b ) DNA, c, d HypaCas9 with on-target ( c ) and off-target ( d ) DNA. e, f Cas9-HF1 with on-target ( e ) and off-target ( f ) DNA. g Cartoon of the <t>HNH</t> domain rearrangement. h The <t>FRET</t> SpCas9 labeled with Cy3 and Cy5 pair at C867 and C355, respectively, was used to measure the HNH domain motion during cleavage of on-target substrates.
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    R-loop formation decay rates were measured by the monitoring the fluorescence increase from tC o (position −1 in the non-target strand) as a function of time after mixing enzyme (28 nM) with DNA (10 nM) in the presence of 10 mM Mg 2+ . Data were fit to a double exponential function to get the decay rates shown. a, b SpCas9 (28 nm) with on-target ( a ) and off-target ( b ) DNA, c, d HypaCas9 with on-target ( c ) and off-target ( d ) DNA. e, f Cas9-HF1 with on-target ( e ) and off-target ( f ) DNA. g Cartoon of the HNH domain rearrangement. h The FRET SpCas9 labeled with Cy3 and Cy5 pair at C867 and C355, respectively, was used to measure the HNH domain motion during cleavage of on-target substrates.

    Journal: Nature Communications

    Article Title: Engineered CRISPR/Cas9 enzymes improve discrimination by slowing DNA cleavage to allow release of off-target DNA

    doi: 10.1038/s41467-020-17411-1

    Figure Lengend Snippet: R-loop formation decay rates were measured by the monitoring the fluorescence increase from tC o (position −1 in the non-target strand) as a function of time after mixing enzyme (28 nM) with DNA (10 nM) in the presence of 10 mM Mg 2+ . Data were fit to a double exponential function to get the decay rates shown. a, b SpCas9 (28 nm) with on-target ( a ) and off-target ( b ) DNA, c, d HypaCas9 with on-target ( c ) and off-target ( d ) DNA. e, f Cas9-HF1 with on-target ( e ) and off-target ( f ) DNA. g Cartoon of the HNH domain rearrangement. h The FRET SpCas9 labeled with Cy3 and Cy5 pair at C867 and C355, respectively, was used to measure the HNH domain motion during cleavage of on-target substrates.

    Article Snippet: Plasmid pMJ806 containing Streptococcus pyogenes Cas9 and plasmid pSHS306 containing the HNH FRET variant of Streptococcus pyogenes Cas9 were obtained from Addgene (Cambridge, MA).

    Techniques: Fluorescence, Labeling