Custom Lentiviral sgRNA Expression Construct with Cas9 gene for Single-Vector CRISPR System (plasmid)

Easily get the CRISPR KO, CRISPRi, or CRISPRa sgRNA Constructs you want

Product Description

The CRISPR/Cas9 system can be used for gene knockout (KO), knockdown, activation or to initiate knock-ins in vivo or in vitro by using a combination of an sgRNA (single-guide RNA) together with a Cas9 nuclease. Some examples of custom constructs include:

• Constitutive or tet-inducible sgRNA constructs designed for CRISPR KO, CRISPRa, or CRISPRi.
• All-in-one constructs with sgRNA and Cas9 or single-vector sgRNA-only formats.
• Complete panels of Cas9 and dCas9-hybrid (e.g., dCas9-KRAB, dCas9-VPH, dCas9-VPR, etc.) expression constructs

Cellecta's improved sgRNA design enables better CRISPR Knockout Screen results. Learn more about the HEAT-Modified sgRNA Design.

We offer both, a Two-Vector System using sgRNA-only constructs together with Cas9 or dCas9 variants on a separate construct and a Single-Vector System with an sgRNA-Cas9 construct expressing both the Cas9 gene and your sgRNA (example see Fig.1). Just provide the gene information and Cellecta will select the optimal guide designs.

Fig.1. Popular All-in-One sgRNA cloning vector pRSGCCP-(sg)-CMV-Cas9-2A-Puro, 11,505 bp

Related Links

CRISPRi and CRISPRa: Beyond Gene Knockout

Performance Data

CRISPR sgRNA constructs targeting GFP were built using the Two Vector and Single Vector CRISPR systems and used to transduce cells that had been previously engineered to stably express GFP. Both CRISPR systems provide at least 80% knockout of GFP, but the two-vector system performed better with over 90% knockout. The enhanced performance is due to pre-selection of cells expressing high levels of Cas9 before transduction with the GFP sgRNA constructs.

Fig.2. Several sgRNAs targeting the copGFP gene were designed and individually transduced into HEK293 cells stably expressing copGFP. After 9 days, cells were imaged for GFP fluorescence. For each sgRNA tested, GFP expression was abolished in at least 70% of transduced cells.

Fig.3. After 9 or 14 days. cells were analyzed by FACS. GFP expression was abolished in at least 70% of the analyzed cells after day 9 and at least 90% after day 14.

Fig.4. CRISPR Knockout with tet-inducible sgRNA constructs

Related Services

Lentiviral Packaging Service for Cas9 constructs
Lentiviral Packaging Service for non-Cas9 constructs