Development of Plant Prime-Editing Systems for Precise Genome Editing.
Development of Plant Prime-Editing Systems for Precise Genome Editing.
复制标题
开发用于精确基因组编辑的植物主要编辑系统。
DOI：
10.1016/j.xplc.2020.100043
复制
发表时间：
2020-05-11
影响因子：
10.5
通讯作者：
Wei P
中科院分区：
文献类型：
--
作者：
Xu R;Li J;Liu X;Shan T;Qin R;Wei P
原始编辑系统具有在人类细胞中执行高效和精确的基因组编辑的能力。在这项研究中，我们首先开发了一个植物素编码子2(PPE2)系统，并通过在水稻HPT-ATG报告基因上产生靶向突变来测试其活性。我们的结果表明，pPE2系统可以在不同的基因组位置诱导可编程编辑。在转基因T0植株中，pPE2产生突变体的频率为0%-31.3%，这表明pPE2的效率在不同的基因组位置和不同结构的初级编辑引导RNA之间存在很大差异。为了优化编辑效率，遵循人类细胞中的PE3和PE3b策略，将引导RNA引入pPE2系统。然而，在这项研究中测试的基因组位置，pPE3系统只产生了类似的甚至更低的编辑频率。此外，我们还开发了一个替代pPE2系统，通过引入HPT-ATG报告基因来丰富原始编辑的细胞。在用替代pPE2系统转化的抗性愈伤组织中很容易检测到核苷酸编辑，这可能是因为编辑细胞的筛选效率提高了。综上所述，我们的结果表明，我们开发的植物素材编辑系统可以提供对水稻基因组的通用和灵活的编辑。本研究开发了植物素编辑(PE)系统，包括pPE2、pPE3/3b和代理pPE2，并验证了该系统能够在转基因水稻中实现精确和通用的基因组编辑。这些PE系统为植物基因组编辑提供了有用的工具。
Prime-editing systems have the capability to perform efficient and precise genome editing in human cells. In this study, we first developed a plant prime editor 2 (pPE2) system and test its activity by generating a targeted mutation on an HPT-ATG reporter in rice. Our results showed that the pPE2 system could induce programmable editing at different genome sites. In transgenic T0 plants, pPE2-generated mutants occurred with 0%–31.3% frequency, suggesting that the efficiency of pPE2 varied greatly at different genomic sites and with prime-editing guide RNAs of diverse structures. To optimize editing efficiency, guide RNAs were introduced into the pPE2 system following the PE3 and PE3b strategy in human cells. However, at the genomic sites tested in this study, pPE3 systems generated only comparable or even lower editing frequencies. Furthemore, we developed a surrogate pPE2 system by incorporating the HPT-ATG reporter to enrich the prime-edited cells. The nucleotide editing was easily detected in the resistant calli transformed with the surrogate pPE2 system, presumably due to the enhanced screening efficiency of edited cells. Taken together, our results indicate that plant prime-editing systems we developed could provide versatile and flexible editing in rice genome. Plant prime-editing (PE) systems, including pPE2, pPE3/3b, and surrogate pPE2, were developed in this study and validated to mediate precise and versatile genome editing in transgenic rice plants. These PE systems provide useful tools for genome editing in plants.
登录
查看更多内容
影响因子：
64.8
作者：
Komor AC;Kim YB;Packer MS;Zuris JA;Liu DR
通讯作者：
Liu DR
影响因子：
64.8
作者：
Anzalone, Andrew V.;Randolph, Peyton B.;Liu, David R.
通讯作者：
Liu, David R.
影响因子：
12.3
作者：
Li C;Zong Y;Wang Y;Jin S;Zhang D;Song Q;Zhang R;Gao C
通讯作者：
Gao C
通过修改水稻胞苷碱基编辑系统来提高保真度和效率
影响因子：
6.6
作者：
Qin, Ruiying;Liao, Shengxiang;Wei, Pengcheng
通讯作者：
Wei, Pengcheng
非等位基因乙酰乳酸合酶基因的精确碱基编辑赋予玉米磺酰脲类除草剂抗性
影响因子：
6.6
作者：
Li, Yanmin;Zhu, Jinjie;Xie, Chuanxiao
通讯作者：
Xie, Chuanxiao