Overview

Enhance your All-in-one Cas9 SmartNickase Plasmid with multiplexed gRNA delivery

Easily enhance the power of the CMV-T7-hspCas9-Nickase-H1-gRNA All-in-one Cas9 SmartNickase™ Plasmid by combining it with the Multiplex gRNA Cloning Kit. Together, these two products can enable a wider range of genome editing projects that require multiple gRNAs.

You get all the great features of the CMV-T7-hspCas9-Nickase-H1-gRNA All-in-one Cas9 SmartNickase Plasmid:

• Conveniently deliver Cas9 and gRNA with a single vector
• Reduce off-target activity
• Drive Cas9 expression with the CMV promoter, which provides high expression levels in many common cell lines (HeLa, HEK293, HT1080)
• 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

Combined with the advantages of the Multiplex gRNA Cloning Kit:

• Edit multiple loci at once, save time and reagents
• Easily generate multi-cistronic gRNA expression constructs
• Ideal for Cas9 Nickase applications
• Great for studying signaling pathways

As with all of our Cas9 delivery options, the CMV-T7-hspCas9-Nickase-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.

Not sure whether you need a CRISPR/Cas9 plasmid, purified protein, or mRNA?

Use this table to choose the CRISPR/Cas9 product that’s right for you: