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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 CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA) for genomic DNA target recognition and cleavage using two active sites that together generate DNA double-strand breaks (DSB).

    Choose from a variety of Cas9 expressing vectors

    The Edit-R Cas9 nuclease expression vectors incorporate a human codon-optimized version of the Streptococcus pyogenes Type II Cas9 (formerly csn1) gene. The activity of any given promoter controlling the transcription of Cas9 nuclease can differ greatly from one biological context to another, resulting in variable Cas9 expression levels and thus varying levels of DNA cleavage. Vector-based Edit-R Cas9 Nuclease products are offered with up to 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 format, thus 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

    Transient Cas9 products enable a completely DNA-free knockout experiment

    DNA-free Cas9 nuclease options protect against long-term expression of Cas9 nuclease when transient expression is desired. With Cas9 mRNA and Cas9 protein there is also no risk of plasmid integration into the host genome or side-effects from constitutive Cas9 expression.

    Vector-based Cas9 Nuclease

    DNA-free Cas9 Nuclease

    Cas9 Nuclease Selection Guide

    While the best Cas9 nuclease product for your experiment may heavily depend on the particular application or cell type, a few basic questions may help to point you in the right direction for product selection.

    Cas9 Nuclease Selection GuideEdit-R Cas9 Lentiviral Plasmid DNAEdit-R Cas9 Lentiviral ParticlesCas9 Nuclease mRNA or proteinEdit-R Cas9 Expression Plasmid

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