Helper Virus-Free Preparation of AAV particles to deliver Cas9 and sgRNA
The AAVpro CRISPR/Cas9 Helper Free Systems are used for helper virus-free preparation of AAV2 particles to deliver expression cassettes for Cas9 and a user-defined single guide RNA (sgRNA) to mammalian cells. This two-vector system overcomes the size restrictions of the AAV genome to deliver the large Cas9 gene to target cells, allowing for efficient genome modification in a wide variety of mammalian cells in vitro. High-titer AAV2 particles can be prepared without a helper virus by simply transfecting HEK 293T cells with the included plasmids that encode all of the factors necessary to prepare recombinant AAV particles. The AAVpro CRISPR/Cas9 Helper Free System is a complete system, containing all the reagents necessary to clone the sgRNA into the expression plasmid and prepare AAV particles. AAV Extraction Solution is also included for efficient AAV2 viral particle isolation.