Osadchiy I S, Kamalyan S O, Tumashova K Y, Georgiev P G, Maksimenko O G
Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334 Russian Federation.
Acta Naturae. 2023 Apr-Jun;15(2):70-74. doi: 10.32607/actanaturae.11874.
Since the addition of the CRISPR/Cas9 technology to the genetic engineering toolbox, the problems of low efficiency and off-target effects hamper its widespread use in all fields of life sciences. Furthermore, essential gene knockout usually results in failure and it is often not obvious whether the gene of interest is an essential one. Here, we report on a new strategy to improve the CRISPR/Cas9 genome editing, which is based on the idea that editing efficiency is tightly linked to how essential the gene to be modified is. The more essential the gene, the less the efficiency of the editing and the larger the number of off-targets, due to the survivorship bias. Considering this, we generated deletions of three essential genes in : and , using fly strains with previous target gene overexpression ("pre-rescued" genetic background).
自从CRISPR/Cas9技术被添加到基因工程工具库以来，效率低下和脱靶效应问题阻碍了其在生命科学所有领域的广泛应用。此外，必需基因敲除通常会导致失败，而且感兴趣的基因是否为必需基因往往并不明显。在此，我们报告一种改进CRISPR/Cas9基因组编辑的新策略，该策略基于编辑效率与待修饰基因的必需程度紧密相关的观点。由于生存偏差，基因越必需，编辑效率越低，脱靶数量就越多。考虑到这一点，我们使用先前靶基因过表达的果蝇品系（“预拯救”遗传背景），在[具体物种或实验对象]中产生了三个必需基因的缺失。
