Chew, Wei LeongTabebordbar, MohammadsharifCheng, Jason K WMali, PrashantWu, Elizabeth YNg, Hon Man AlexZhu, KexianWagers, AmyChurch, George2022-07-192016-09-05Chew, Wei Leong, Mohammadsharif Tabebordbar, Jason K W Cheng, Prashant Mali, Elizabeth Y Wu, Hon Man Alex Ng, Kexian Zhu et al. "A Multifunctional AAV–CRISPR–Cas9 and Its Host Response." Nature Methods 13, no. 10 (2016): 868-874. DOI: 10.1038/nmeth.39931548-70911548-7105https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37372659CRISPR-Cas9 delivery by AAV holds promise for gene therapy but faces critical barriers due to its potential immunogenicity and limited payload capacity. Here, we demonstrate genome engineering in postnatal mice using AAV-split-Cas9, a multi-functional platform customizable for genome-editing, transcriptional regulation, and other previously impracticable AAV-CRISPR-Cas9 applications. We identify crucial parameters that impact efficacy and clinical translation of our platform, including viral biodistribution, editing efficiencies in various organs, antigenicity, immunological reactions, and physiological outcomes. These results reveal that AAV-CRISPR-Cas9 evokes host responses with distinct cellular and molecular signatures, but unlike alternative delivery methods, does not induce extensive cellular damage in vivo. Our study provides a foundation for developing effective genome therapeutics.en-USResearch Subject Categories::TECHNOLOGY::Bioengineering::Genetic engineering including functional genomicsResearch Subject Categories::MEDICINE::Physiology and pharmacology::Physiology::Laboratory animal scienceJournal Article2017-08-0220162022-07-1910.1038/nmeth.3993