Cas9 Lentivirus (Neomycin Selection)

Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease enzyme that, when recruited to a specific DNA sequence by the sgRNA (single guide RNA), introduces a double stranded break into the DNA. This double stranded break is repaired in mammalian cells either through Non-Homologous End Joining or Homologous Recombination. Non-Homologous End Joining often results in the deletion or insertion of several base pairs at the cut site, which, when resulting in a frameshift, causes the functional inactivation of the targeted gene. Cas9 Lentivirus can be used to generate Cas9 expressing cells in almost any mammalian cell line. Cells stably expressing Cas9 can then be transduced or electroporated with sgRNA targeting a gene of interest to quickly generate knock-out cell pools or cell lines. The Cas9 Lentiviruses are replication incompetent, HIV-based VSV-G pseudo-typed lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain a Cas9 gene driven by an EF1a promoter, along with a Neomycin selection marker.