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dcas9 krab p2a mcherry cassette  (Addgene inc)


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    Addgene inc dcas9 krab p2a mcherry cassette
    Dcas9 Krab P2a Mcherry Cassette, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dcas9+krab+p2a+mcherry+cassette/pHR-SFFV-KRAB-dCas9-P2A-mCherry+(Plasmid+%2360954)/pm41832176-299-1-7
    Average 95 stars, based on 128 article reviews
    dcas9 krab p2a mcherry cassette - by Bioz Stars, 2026-10
    95/100 stars

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    Plasmid Preparation:

    Article Title: Cell type-specific enhancers regulate IL-22 expression in innate and adaptive lymphoid cells
    Article Snippet: Next, TRE3G promoter and pHR-pEF-EGFP vector (Addgene Plasmid #67952) were digested with EcoR1+BamH1 and ligated, resulting in pHR-TRE3G-EGFP vector. .. Lastly, dCas9-KRAB-P2A-mcherry cassette was swapped from pHR-SFFV-KRAB-dCas9-P2A-mCherry (Addgene Plasmid #60954) into pHR-TRE3G-EGFP vector from above using Mlu1 and Not1 restriction sites. sgRNAs targeting Il22 promoter, E22-1, E22-2, twin-E22-2, or scrambled sgRNA were cloned downstream of the U6 promoter in sgOpti (Addgene Plasmid #85681). ..

    Article Title: Cell type-specific enhancers regulate IL-22 expression in innate and adaptive type 3 lymphoid cells.
    Article Snippet: Next, TRE3G promoter and pHR-pEFEGFP vector47 (Addgene Plasmid #67952) were digested with EcoR1+BamH1 and ligated, resulting in pHR-TRE3G-EGFP vector. .. Lastly, dCas9-KRAB-P2A-mcherry cassette was swapped from pHR-SFFVKRAB-dCas9-P2A-mCherry48 (Addgene Plasmid #60954) into pHR-TRE3G-EGFP vector from above using Mlu1 and Not1 restriction sites. sgRNAs targeting Il22 promoter, E22-1, E22-2, twin-E22-2, or scrambled sgRNA were cloned downstream of the U6 promoter in sgOpti49 (Addgene Plasmid #85681). ..

    Clone Assay:

    Article Title: Cell type-specific enhancers regulate IL-22 expression in innate and adaptive lymphoid cells
    Article Snippet: Next, TRE3G promoter and pHR-pEF-EGFP vector (Addgene Plasmid #67952) were digested with EcoR1+BamH1 and ligated, resulting in pHR-TRE3G-EGFP vector. .. Lastly, dCas9-KRAB-P2A-mcherry cassette was swapped from pHR-SFFV-KRAB-dCas9-P2A-mCherry (Addgene Plasmid #60954) into pHR-TRE3G-EGFP vector from above using Mlu1 and Not1 restriction sites. sgRNAs targeting Il22 promoter, E22-1, E22-2, twin-E22-2, or scrambled sgRNA were cloned downstream of the U6 promoter in sgOpti (Addgene Plasmid #85681). ..

    Article Title: Cell type-specific enhancers regulate IL-22 expression in innate and adaptive type 3 lymphoid cells.
    Article Snippet: Next, TRE3G promoter and pHR-pEFEGFP vector47 (Addgene Plasmid #67952) were digested with EcoR1+BamH1 and ligated, resulting in pHR-TRE3G-EGFP vector. .. Lastly, dCas9-KRAB-P2A-mcherry cassette was swapped from pHR-SFFVKRAB-dCas9-P2A-mCherry48 (Addgene Plasmid #60954) into pHR-TRE3G-EGFP vector from above using Mlu1 and Not1 restriction sites. sgRNAs targeting Il22 promoter, E22-1, E22-2, twin-E22-2, or scrambled sgRNA were cloned downstream of the U6 promoter in sgOpti49 (Addgene Plasmid #85681). ..



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    Primers, plasmids, and gene block DNA sequences used for BAC16 recombination (5′→3′)
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    Primers, plasmids, and gene block DNA sequences used for BAC16 recombination (5′→3′)

    Journal: Journal of Virology

    Article Title: Rainbow Kaposi's Sarcoma-Associated Herpesvirus Revealed Heterogenic Replication with Dynamic Gene Expression

    doi: 10.1128/JVI.01565-19

    Figure Lengend Snippet: Primers, plasmids, and gene block DNA sequences used for BAC16 recombination (5′→3′)

    Article Snippet: mCherry cassette template, Addgene 60954 , .

    Techniques: Blocking Assay, Plasmid Preparation, Sequencing

    Schematic diagram of procedures to prepare Rainbow-KSHV. (i) The cDNA of fluorescence tag coding sequence (mBFP2, mCardinal, or mCherry) first was cloned into pBS vector. (ii and iii) The kanamycin cassette with I-SceI recognition sequence along with 50-bp homology sequence was amplified by PCR and cloned into the indicated unique restriction enzyme site (AccI or PstI). (iv and v) The resulting plasmid was used as a template to generate a DNA fragment for homologous recombination with BAC16 inside bacteria. (vi and vii) After confirmation of insertion at the correct site by colony PCR screening, the kanamycin cassette was deleted by recombination after induction of I-SceI in bacteria by l -arabinose. Correct insertion of the fluorescence tags and integrity of BAC DNA were confirmed by sequencing of PCR-amplified fragments and restriction digestions. (viii and ix) Additional viral proteins were similarly tagged with mCherry and mCardinal fluorescent protein. Primers and DNA fragment used are listed in .

    Journal: Journal of Virology

    Article Title: Rainbow Kaposi's Sarcoma-Associated Herpesvirus Revealed Heterogenic Replication with Dynamic Gene Expression

    doi: 10.1128/JVI.01565-19

    Figure Lengend Snippet: Schematic diagram of procedures to prepare Rainbow-KSHV. (i) The cDNA of fluorescence tag coding sequence (mBFP2, mCardinal, or mCherry) first was cloned into pBS vector. (ii and iii) The kanamycin cassette with I-SceI recognition sequence along with 50-bp homology sequence was amplified by PCR and cloned into the indicated unique restriction enzyme site (AccI or PstI). (iv and v) The resulting plasmid was used as a template to generate a DNA fragment for homologous recombination with BAC16 inside bacteria. (vi and vii) After confirmation of insertion at the correct site by colony PCR screening, the kanamycin cassette was deleted by recombination after induction of I-SceI in bacteria by l -arabinose. Correct insertion of the fluorescence tags and integrity of BAC DNA were confirmed by sequencing of PCR-amplified fragments and restriction digestions. (viii and ix) Additional viral proteins were similarly tagged with mCherry and mCardinal fluorescent protein. Primers and DNA fragment used are listed in .

    Article Snippet: mCherry cassette template, Addgene 60954 , .

    Techniques: Fluorescence, Sequencing, Clone Assay, Plasmid Preparation, Amplification, Homologous Recombination, Bacteria