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Applications: Generation of Tet-inducible Cas9 stable cell lines (Geneticin (G418) selection) for genetic engineering studies.
Background: Cas9 (Streptococcus pyogenes CRISPR associated protein 9) is an endonuclease that, when recruited to a specific DNA sequence by the appropriate sgRNA (single guide RNA), introduces a double stranded break into the DNA. This double stranded break can then be repaired through either Non-Homologous End Joining (NHEJ) or Homologous Recombination (HR). While NHEJ is an error prone process and causes insertions or deletions which may result in functional inactivation of the target gene, HR, in conjunction with a single stranded ssDNA repair construct, can be used to introduce mutations at specific base pair(s). Gene modifications introduced via Cas9 are now used in multiple fields of research aimed at understanding cellular mechanisms and developing therapeutic solutions.
Description: Cas9 Lentiviruses (Inducible Tet-On) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses transduce cells with the Streptococcus pyogenes Cas9 gene under a tight TRE tetracycline-inducible promoter. Cas9 expression in the transduced cells is induced with doxycycline treatment, allowing temporal control of its expression, and when combined with sgRNA targeting gene(s) of interest allows gene editing events to be temporally controlled. The lentivirus vector also contains a geneticin selection gene.
Formulation: The lentiviruses were produced from HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.
Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the lentiviruses at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
Supplied As: Two vials (500 µl x 2) of lentivirus at a titer 1 x 107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-2
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