We offer two kits for helper virus-free preparation of AAV2 particles for genome editing with Cas9: the AAVpro CRISPR/Cas9 Helper Free System and the AAVpro CRISPR/SaCas9 Helper Free System. These kits simultaneously deliver expression cassettes for Cas9 and a user-defined single guide RNA (sgRNA) to mammalian cells. Cas9 sequences are derived from either Streptococcus pyogenes (the CRISPR/Cas9 systems) or Staphylococcus aureus (the CRISPR/SaCas9 systems).

The use of an AAV system allows for efficient genome modification in a wide variety of mammalian cells in vitro, with the CRISPR/Cas9 system using two AAV vectors to deliver the larger S. pyogenes Cas9 (SpCas9), while the CRISPR/SaCas9 system delivers everything in a single vector due to the smaller size of the SaCas9 gene. sgRNAs can be cloned directly into the prelinearized pAAV-Guide-it-Down or pAAV-Guide-it-1 plasmids in a single step. High-titer AAV2 particles can be prepared without a helper virus by simply transfecting HEK 293T cells with the included plasmids. Both the AAVpro CRISPR/Cas9 Helper Free System (Cat. # 632608) and the AAVpro CRISPR/SaCas9 Helper Free System (Cat. # 632619) are complete systems, containing all the reagents necessary to clone sgRNAs and prepare AAV particles. AAV Extraction Solution is also included for efficient AAV2 viral particle isolation. Please note that SpCas9 and SaCas9 can have different targeting efficiencies even when used on the same region of the genome—refer to Image Data (via the Details arrow in the product table below) for more information.