The mammalian zygote-mediated CRISPR/Cas system can efficiently generate targeted genome-modified animals. However, this system is limited by the risk of off-target mutations. Here we show that offset-nicking by Cas9 nickase and paired gRNAs allows us to generate region deleted mice and targeted knock-in mice without off-target mutations.
Keywords: CRISPR/Cas9; Genome-modified animals; Nickase; Off-target.
Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.