Off-Target Effects Of CRISPR/Cas9 and Their Solutions

Abstract

In the continuous exploration, the superiority of CRISPR/Cas9 system has been discovered. Its ability to edit target genes with high specific and low cost has greatly increased the convenience of constructing biological models, accelerated the screening of targets and effectively improve the gene editing technology. However, when CRISPR/Cas9 system is used in the studies, these studies are frequently constrained by the off-target effects, which may result in mutations in the experimental products and erroneous experimental data. Therefore, this paper mainly introduces CRISPR/Cas9 and the principle of its off-target, and discusses the effects of off-target from PAM sequence, sgRNA and other aspects. To more thoroughly study the off-target effects' mechanism, this paper also lists three kinds of detection technologies of off-target effects. Finally, three solutions including designing sgRNA, changing the abundance of Cas9/sgRNA and improving Cas9 protein were analyzed and discussed from the effects of off-target, which provided ideas for reducing the off-target effects and improving genome editing technology.