AAVpro CRISPR/Cas9 Systems
- CRISPR/CAS9 genome editing in hard-to-transfect mammalian cells
- Simultaneously deliver Cas9 and sgRNA to cells
- Non-integrating virus: avoids Cas9 off-target effects
- Prelinearized vectors facilitate single-step sgRNA cloning
- Contains all reagents from sgRNA cloning to AAV particle preparation
- No need for a helper virus to produce the AAV2 particles
With the AAVpro CRISPR/Cas9 System you can prepare adeno-associated virus (AAV) particles needed for CRISPR/Cas9-mediated genome editing, without the need of a helper virus. It can be used for a wide variety of mammalian cells.
Deliver Cas9 and sgRNA to your cells without a helper virus
The AAVpro CRISPR/Cas9 Vector Systems consists of two vectors: the pAAV Guide-it-up and pAAV Guide-it-down vectors. Due to this two-vector system, size restrictions of the AAV genome to deliver the large Cas9 gene to target cells can be overcome. The AAVpro CRISPR/Cas9 Helper Free System also includes the human microRNA miR-342 packaging plasmid. Expression of this plasmid significantly increases AAV titers. Below you will find a schematic representation of the AAVpro CRISPR/Cas9 Vector Systems workflow.
Simple 5-step workflow to deliver Cas9/sgRNA to your cells
- Clone your sgRNA into the pAAV-Guide-it-Down vector.
- Transfect both vectors seperately in HEK293 cells. You can increase AAV titers by including the pRC2-mi342 Vector with its pHelper Vector during transfections.
- Isolate the AAV2 particles from both transfections seperately. You can use the AAVpro Extraction Solution to get optimal particle yields.
- Perform a co-tranduction on your target cells with both viruses: tranduce your cells with both viruses in 1 dish. During the transduction, recombination occurs at the region of homology to create a full-length Cas9 gene with an upstream CMV promoter.
- Finally, you can determine AAV titers (you can use the AAVpro AAV Titration kit for this) and the expression of Cas9/sgRNA in your target cells.