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The CRISPR/Cas9 system uses the Cas9 nuclease to facilitate RNA-guided site-specific DNA cleavage. Our system consists of two components:

(1) A mammalian codon-optimized version of the Cas9 protein carrying a nuclear localization signal to ensure nuclear compartmentalization in mammalian cells

(2) Guide RNAs (gRNAs) to direct the Cas9 protein to sequence-specific cleavage the targeted DNA

The advantage of CRISPR/Cas9 over ZFNs or TALENs are scalability and multiplexibility. With CRISPR/Cas9 multiple sites within a mammalian genome can be simultaneously modified, providing a robust, high-throughput approach for gene editing and CRISPR point mutations in a variety of mammalian cells.