Overview

Expanding your genome editing capabilities

Bringing together the versatile CRISPR/Cas9 genome editing system with powerful recombinant AAV (rAAV) technology, SBI’s AAV Cas9 SmartNuclease™ vectors extend genome editing capabilities to cutting edge in vivo applications.

For the most streamlined in vivo introduction of Cas9 using rAAV technology, SBI offers an easy-to-use All-in-one AAV Cas9 SmartNuclease Plasmid EF1α-hsaCas9-U6-gRNA(SA). Simply clone in your gRNA sequence, package the vector, and isolate rAAV virus particles using our AAVanced AAV Concentration Reagent for easy, high-titer preparations.

Please note the “How it Works” section below, which provides instructions for the slightly different gRNA design needed when using hsaCas9.

The saCas9 gene is constitutively driven by the strong EF1α promoter with the gRNA driven by a U6 shRNA promoter.

• Deliver Cas9 in vivo
• Edit genomes in post-natal animals
• Develop gene therapies in small animal models
• Generate novel disease models
• Choose from All-in-one or Two Vector AAV-Cas9 systems

Why AAV?

With their broad tropism, the lack of disease associated with wild-type virus, ability to transduce both dividing and non-dividing cells, and long term transgene expression, recombinant AAV (rAAV) has recently become the method of choice for delivering gene therapy and genome engineering vectors to intact organisms^{1, 2}. However, for efficient packaging, inserts into the region between rAAV’s two ITR sequences must be less than 5 kb.

Why saCas9?

The development of CRISPR/Cas9 has already revolutionized what’s possible when it comes to manipulating the genomes of even complex organisms. However, in vivo delivery via rAAV vectors has been hampered by the size of the Streptococcus pyogenes Cas9 gene (spCas9), the most widely-used form of Cas9. To overcome this problem, Ran, et al,¹ characterized smaller orthologs of the Cas9 gene and found that Cas9 from Staphylococcus aureus (saCas9) performs as efficiently as spCas9 while being ~1 kb shorter, enabling insertion into rAAV vectors.

Why SBI for AAV-Cas9?

With advanced rAAV systems and a range of easy-to-use Cas9 vectors and kits, SBI has the expertise to combine these two technologies into a single, easy-to-use, and powerful system. Choose from our All-in-one or Two Vector systems to drive your in vivo genome editing studies into high gear.

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: