Frequent endonuclease cleavage at off-target locations in vivo.
Target-site DNA breaks increase recombination frequencies, however, the specificity of the enzymes used to create them remains poorly defined. The location and frequency of off-target cleavage events are especially important when rare-cutting endonucleases are used in clinical settings. Here, we identify noncanonical cleavage sites of I-SceI that are frequently cut in the human genome by localizing adeno-associated virus (AAV) vector-chromosome junctions, demonstrating the importance of in vivo characterization of enzyme cleavage specificity.
Petek
Lisa M
LM
Department of Pediatrics, Division of Genetic Medicine, University of Washington, Seattle, Washington, USA.
Russell
David W
DW
Miller
Daniel G
DG
D002460:Cell Line; D053903:DNA Breaks, Double-Stranded; D000229:Dependovirus / Q000235:genetics; D004720:Endonucleases / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans
D004720:Endonucleases