CRISPR Inducible Cas9-BFP Expression Construct pRTCBfpE2Bla-TRE-Cas9BFP-EFS-rtTA-2A-Blast (packaged)

Description

Inducible Lentiviral Cas9 (S. pyogenes) – BFP Fusion Expression Construct

• Fine-tune and time your CRISPR knock-out experiments and screens
• Visualize Cas9 expression and location by blue fluorescence
• Create your own stable cell lines with robustly inducible and tightly regulated Cas9-BFP expression for cell engineering and CRISPR screening
• Ensure that Cas9-BFP 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.

NEW:
We would like to introduce inducible Cas9 BFP Fusion Protein, newly developed by our partner Cellecta.
This inducible all-in-one lentiviral vector is expressing spCas9-BFP 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-BFP. In the off-state there is no Cas9-BFP expression at all. The vector contains all the elements needed for transduction and stable integration of an inducible Cas9-BFP gene for CRISPR-mediated gene editing. The Cas9-BFP 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.