Inducible Lentiviral Cas9 (S. pyogenes) Expression Construct
• Fine-tune and time your CRISPR knock-out experiments and screens
• Create your own stable cell lines with robustly inducible and tightly regulated Cas9 expression for cell engineering and CRISPR screening
• Ensure that Cas9 expression only occurs when it is required
Constitutive expression of Cas9 can result in undesirable outcomes, such as cellular toxicity and off-target effects. This demands generation of inducible CRISPR/Cas9 systems for functional perturbation in a tunable way.
We would like to introduce inducible Cas9, newly developed by our partner Cellecta.
This inducible all-in-one lentiviral vector is expressing spCas9 driven by the Tet-Responsive Promoter (PTRE) and the rtTA tetracycline regulator (Tet-On) and Blast constitutively by EF1alpha. Only in the presence of doxycyclin, the rtTA transactivator protein binds to the pTRE promoter and activates the expression of Cas9. In the off-state there is no Cas9 expression at all. The vector contains all the elements needed for transduction and stable integration of an inducible Cas9 gene for CRISPR-mediated gene editing. The Cas9 protein can be expressed for a limited period to catalyze genomic editing and then shut down for subsequent use of the engineered cells. The vector uses the same optimized hybrid tet-regulated transactivator rtTA that forms the basis of Cellecta’s InDOXible Tet-Activated Lentiviral Expression System.
This construct is also available as plasmid DNA.