We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
High levels of AAV vector integration into CRISPR-induced DNA breaks.
- Authors
Hanlon, Killian S.; Kleinstiver, Benjamin P.; Garcia, Sara P.; Zaborowski, Mikołaj P.; Volak, Adrienn; Spirig, Stefan E.; Muller, Alissa; Sousa, Alexander A.; Tsai, Shengdar Q.; Bengtsson, Niclas E.; Lööv, Camilla; Ingelsson, Martin; Chamberlain, Jeffrey S.; Corey, David P.; Aryee, Martin J.; Joung, J. Keith; Breakefield, Xandra O.; Maguire, Casey A.; György, Bence
- Abstract
Adeno-associated virus (AAV) vectors have shown promising results in preclinical models, but the genomic consequences of transduction with AAV vectors encoding CRISPR-Cas nucleases is still being examined. In this study, we observe high levels of AAV integration (up to 47%) into Cas9-induced double-strand breaks (DSBs) in therapeutically relevant genes in cultured murine neurons, mouse brain, muscle and cochlea. Genome-wide AAV mapping in mouse brain shows no overall increase of AAV integration except at the CRISPR/Cas9 target site. To allow detailed characterization of integration events we engineer a miniature AAV encoding a 465 bp lambda bacteriophage DNA (AAV-λ465), enabling sequencing of the entire integrated vector genome. The integration profile of AAV-465λ in cultured cells display both full-length and fragmented AAV genomes at Cas9 on-target sites. Our data indicate that AAV integration should be recognized as a common outcome for applications that utilize AAV for genome editing. In-depth characterization of adeno-associated virus (AAV)-mediated CRISPR delivery is still lacking. Here, the authors show high levels of integration into Cas9-induced double-strand breaks (DSBs) in therapeutically relevant genes in vivo.
- Subjects
BACTERIOPHAGE lambda; ADENO-associated virus; BRAIN mapping; ANIMAL models in research; GENOME editing; DNA
- Publication
Nature Communications, 2019, Vol 10, Issue 1, pN.PAG
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-019-12449-2