Malzahn Aimee, Lowder Levi, Qi Yiping
Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742 USA.
Department of Biology, East Carolina University, Greenville, NC 27858 USA.
Cell Biosci. 2017 Apr 24;7:21. doi: 10.1186/s13578-017-0148-4. eCollection 2017.
Genome editing promises giant leaps forward in advancing biotechnology, agriculture, and basic research. The process relies on the use of sequence specific nucleases (SSNs) to make DNA double stranded breaks at user defined genomic loci, which are subsequently repaired by two main DNA repair pathways: non-homologous end joining (NHEJ) and homology directed repair (HDR). NHEJ can result in frameshift mutations that often create genetic knockouts. These knockout lines are useful for functional and reverse genetic studies but also have applications in agriculture. HDR has a variety of applications as it can be used for gene replacement, gene stacking, and for creating various fusion proteins. In recent years, transcription activator-like effector nucleases and clustered regularly interspaced palindromic repeats (CRISPR) and CRISPR associated protein 9 or CRISPR from Prevotella and Francisella 1 have emerged as the preferred SSNs for research purposes. Here, we review their applications in plant research, discuss current limitations, and predict future research directions in plant genome editing.
基因组编辑有望在推进生物技术、农业和基础研究方面取得巨大飞跃。该过程依赖于使用序列特异性核酸酶(SSN)在用户定义的基因组位点产生DNA双链断裂,随后通过两种主要的DNA修复途径进行修复:非同源末端连接(NHEJ)和同源定向修复(HDR)。NHEJ可导致移码突变,常产生基因敲除。这些敲除系对于功能和反向遗传学研究很有用,但在农业中也有应用。HDR有多种应用,因为它可用于基因替换、基因堆叠以及创建各种融合蛋白。近年来,转录激活样效应核酸酶和成簇规律间隔短回文重复序列(CRISPR)以及CRISPR相关蛋白9或普氏菌和弗朗西斯菌1型CRISPR已成为研究目的首选SSN。在此,我们综述它们在植物研究中的应用,讨论当前的局限性,并预测植物基因组编辑的未来研究方向。