...into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb). While this approach has...previously packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity...preferential infection of certain tissues. However, packaging Streptococcus pyogenes (SpCas9) and a gRNA together... the only CRISPR enzyme that’s small enough to package into AAV.
At 984 amino acids in length, Cas9 from...Cas9 (NmeCas9) at 1,082 amino acids, can also be packaged into AAV. NmeCas9 has the added advantage that...respectively, the size of both is within AAV’s packaging limit.
For CRISPR-based RNA editing, the REPAIR...approximately 4.1 kb in length, allowing them to be packaged in AAV.
Split AAV approaches
While finding...