Model EF1a-T7-hspCas9-T2A-RFP-H1-gRNA - All-in-one Cas9 SmartNuclease™ Plasmid

Conveniently check transfection efficiencies with this All-in-one Cas9 SmartNuclease

Our All-in-one Cas9 and gRNA plasmids are an excellent way to simplify delivery of your CRISPR/Cas9 system by providing both Cas9 and gRNA from a single vector, and the addition of coordinate expression of RFP for monitoring transfection efficiencies helps make genome engineering projects more user-friendly. SBI’s EF1α-T7-hspCas9-T2A-RFP-H1-gRNA All-in-one Cas9 SmartNuclease Plasmid includes a number of additional features that make it a great All-in-one choice for any genome engineering project involving transfectable cells:

• Conveniently deliver Cas9 and gRNA with a single vector
• Monitor transfection efficiencies with RFP, which is coordinately expressed with the hspCas9 gene via the T2A element
• Drive Cas9 expression with the EF1α promoter, which provides medium expression levels in most cell types, including primary cells and stem cells
• Express gRNA from the H1 promoter for maximum specificity and choice of targets
• Ensure efficient import of Cas9 to the nucleus with N-term and C-term nuclear localization signals (NLSs)
• Boost Cas9 gene expression and stabilize the transcript via the WPRE regulatory element after the C-term NLS
• Easily detect and/or purify the Cas9 protein with the N-term myc-tag
• Produce Cas9 mRNA via in vitro transcription using the T7 promoter

As with all of our Cas9 delivery options, the EF1α-T7-hspCas9-T2A-RFP-H1-gRNA Plasmid is functionally validated and comes backed by our expert technical support team—if you’ve got a genome engineering question just ask by emailing tech@systembio.com.

Why an HR targeting vector is a recommended

Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation. HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events. In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5’ or 3’ homology arms, can make specific, targeted gene edits.

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