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    Cas9 Nuclease

    Configurable expression constructs or DNA-free options for optimal Cas9 expression

    Active expression of the Cas9 endonuclease is a critical requirement for efficient gene editing. The Edit-R portfolio of Cas9 nuclease expression vectors includes six SMARTchoice constitutive promoter options or an inducible format, providing the flexibility to select an optimal system for robust expression and maximal cleavage efficiency when Edit-R guide RNAs are introduced into the cell.

    Cas9 nuclease in the CRISPR-Cas9 system

    In Type II CRISPR-Cas systems, the CRISPR-associated enzyme Cas9 is an RNA-guided endonuclease that requires a guide RNA for genomic DNA target recognition and cleavage. All Edit-R Cas9 nuclease products are a human codon-optimized version of the Streptococcus pyogenes Type II Cas9 nuclease.

    Fluorescent Cas9 reagents for optimization and enrichment

    Enrichment of edited cells in a population is greatly simplified with Cas9 fluorescent lentiviral or mRNA reagents that co-express either mKate2 or TurboGFP™ with Cas9. These reagents also allow for straightforward transfection optimization and visualization of successful Cas9 delivery.

    Transient Cas9 products enable a completely DNA-free knockout experiment

    Cas9 nuclease mRNA and protein are DNA-free options that permit transient expression of Cas9 to reduce off-targets and protect cells from potential unwanted cellular responses from long term expression. Fluorescent Cas9 mRNA allows for sorting, enrichment and visualization of Cas9 delivery.

    Choose from a variety of Cas9 vectors for optimal expression in your system

    Promoter activity, and resultant Cas9 expression, can vary with cell type; therefore vector-based Edit-R Cas9 Nuclease products are offered with six different, well-characterized, constitutive cellular promoters so you can choose an optimal promoter for your cells (Table 1). Edit-R Cas9 Nuclease is also available with an inducible promoter, permitting you to control the timing of the expression of Cas9, which may be a critical component to a successful experiment.

    Table 1. Six SMARTchoice promoter options for expressing Cas9 nuclease
    Promoter Description
    hCMV human cytomegalovirus immediate early promoter
    mCMV mouse cytomegalovirus immediate early promoter
    hEF1α human elongation factor 1 alpha promoter
    mEF1α mouse elongation factor 1 alpha promoter
    PGK mouse phosphoglycerate kinase promoter
    CAG chicken beta actin hybrid promoter
    TRE3G doxycycline inducible promoter

    Vector-based Cas9 Nuclease

    • Edit-R Cas9 Nuclease Expression reagents

      Non-lentiviral vectors provided as endotoxin-free purified DNA for direct co-transfection with Edit-R synthetic single guide RNA or crRNA and tracrRNA.

    • Edit-R Lentiviral Cas9 Nuclease reagents

      Purified lentiviral particles or plasmid DNA for generation of stable Cas9 nuclease-expressing cell populations. Constitutive or inducible promoter options are available.

    DNA-free Cas9 Nuclease

    Cas9 Nuclease Selection Guide

    Cas9 nuclease is not one-size-fits-all. Determining the most appropriate Cas9 nuclease reagent for your experiment is dependent on the particular application or cell type. Take a look at the features in this table to get started on selecting the best product for you.