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    Edit-R dCas9-VPR

    The CRISPR activation (CRISPRa) system is an adapted CRISPR-Cas9 system that is used for upregulation of genes. It utilizes a nuclease-deactivated S. pyogenes Cas9 (dCas9), that is fused to transcriptional activation domains. In the VPR activation system, dCas9 is fused at the C-terminal end to three transcriptional activators (VP64, p65 and Rta). When paired with guide RNAs that target a gene near a promoter region, the gene's native transcription start site is activated.

    There are several format options for dCas9-VPR. The Edit-R dCas9-VPR mRNA allows for direct co-transfection or electroporation of reagents and provides options for enrichment by fluorescence or antibiotic selection. Edit-R Lentiviral dCas9-VPR reagents can be used to generate a stable population of dCas9-VPR cells which is ideal for screening and for extended timepoint assays. Edit-R Lentiviral dCas9-VPR reagents are available with three different promoters (hCMV, mCMV, or hEF1a) and can be supplied as either purified high-titer lentiviral particles or purified plasmid DNA.

    CRISPR-Cas9 for gene activation

    CRISPR-Cas9 is not only for creating genomic double-strand breaks, it can also be used to target promoter regions to activate or inhibit transcription. Using a deactivated or dead Cas9 nuclease fused to transcription activators, like Cas9-VPR, in combination with a guide RNA in either a synthetic or lentiviral format, the endogenous expression of a gene can be up-regulated by anywhere from 5 to 50,000+ fold!

    dCas9-VPR reagents for optimization and enrichment

    Enrichment for gene activation can now easily be done using the dCas9-VPR mRNA reagents that co-express either EGFP or puromycin with dCas9-VPR. These reagents allow easy co-transfection or electroporation for optimization and visualization of successful delivery.

    Choose from a variety of dCas9-VPR vectors

    Promoter activity will vary by cell type and will affect dCas9-VPR expression, choosing an optimal promoter is important for robust gene overexpression. Edit-R Lentiviral dCas9-VPR reagents are available with three different promoters (hCMV, mCMV, or hEF1a) and can be supplied as either purified high-titer lentiviral particles or purified plasmid DNA.

    Table 1. SMARTchoice promoter options for expressing dCas9-VPR nuclease
    Promoter Description
    hCMV human cytomegalovirus immediate early promoter
    mCMV mouse cytomegalovirus immediate early promoter
    hEF1α human elongation factor 1 alpha promoter

    DNA-free dCas9-VPR

    • Edit-R dCas9-VPR mRNA

      Purified dCas9-VPR mRNA for transient expression. Fluorescent and puromycin options available for sorting, enrichment, and visualization of delivery

    Vector-based dCas9-VPR