Guha Tuhin Kumar, Wai Alvan, Hausner Georg
Department of Microbiology, University of Manitoba, Winnipeg, Manitoba R3T2N2, Canada.
Comput Struct Biotechnol J. 2017 Jan 12;15:146-160. doi: 10.1016/j.csbj.2016.12.006. eCollection 2017.
Targeted genome editing has become a powerful genetic tool for studying gene function or for modifying genomes by correcting defective genes or introducing genes. A variety of reagents have been developed in recent years that can generate targeted double-stranded DNA cuts which can be repaired by the error-prone, non-homologous end joining repair system or via the homologous recombination-based double-strand break repair pathway provided a suitable template is available. These genome editing reagents require components for recognizing a specific DNA target site and for DNA-cleavage that generates the double-stranded break. In order to reduce potential toxic effects of genome editing reagents, it might be desirable to control the in vitro or in vivo activity of these reagents by incorporating regulatory switches that can reduce off-target activities and/or allow for these reagents to be turned on or off. This review will outline the various genome editing tools that are currently available and describe the strategies that have so far been employed for regulating these editing reagents. In addition, this review will examine potential regulatory switches/strategies that can be employed in the future in order to provide temporal control for these reagents.
靶向基因组编辑已成为一种强大的遗传学工具，可用于研究基因功能，或通过纠正缺陷基因或引入基因来修饰基因组。近年来已开发出多种试剂，它们能够产生靶向双链DNA切割，这些切割可通过易出错的非同源末端连接修复系统进行修复，或者在有合适模板的情况下，通过基于同源重组的双链断裂修复途径进行修复。这些基因组编辑试剂需要用于识别特定DNA靶位点以及用于产生双链断裂的DNA切割的组件。为了降低基因组编辑试剂的潜在毒性作用，通过引入能够减少脱靶活性和/或允许这些试剂开启或关闭的调控开关来控制这些试剂在体外或体内的活性可能是可取的。本综述将概述目前可用的各种基因组编辑工具，并描述迄今为止用于调控这些编辑试剂的策略。此外，本综述将探讨未来可采用的潜在调控开关/策略，以便为这些试剂提供时间控制。
