Mini-PE是一种具有紧凑型Cas9和截短逆转录酶的主编者。
Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase.
发表日期：2023 Sep 12
作者：
Ting Lan, Huangyao Chen, Chengcheng Tang, Yuhui Wei, Yang Liu, Jizeng Zhou, Zhenpeng Zhuang, Quanjun Zhang, Min Chen, Xiaoqing Zhou, Yue Chi, Jinling Wang, Yu He, Liangxue Lai, Qingjian Zou
来源：
BIOMEDICINE & PHARMACOTHERAPY
摘要：
Prime editor（PE）是一种多功能基因组编辑工具，不需要额外的DNA供体或引发双链断裂。然而，PE在体内的应用仍然具有挑战性，因为其体积过大。在本研究中，我们筛选出具有F155Y突变的Moloney小鼠白血病病毒（MMLV）逆转录酶（RT）的最小截断型，以保持基因编辑效率。我们发现了最高效的MMLV RT变异体，其体积最小。在对pegRNA进行优化并与nick sgRNA结合之后，mini-PE能够在人类和小鼠细胞中以高达10%的准确编辑效果传递到目标位点。尽管编辑效率低于1%，但mini-PE在与腺相关病毒（AAVs）共传递之后，也能够准确地编辑小鼠视网膜中的Hsf1基因。未来，我们将重点改进mini-PE的编辑效率，并开发其对人类遗传疾病的治疗潜力。© 2023 The Author(s).
Prime editor (PE) is a versatile genome editing tool that does not need extra DNA donors or inducing double-strand breaks. However, in vivo implementation of PE remains a challenge because of its oversized composition. In this study, we screened out the smallest truncated Moloney murine leukemia virus (MMLV) reverse transcriptase (RT) with the F155Y mutation to keep gene editing efficiency. We discovered the most efficient gene editing variants of MMLV RT with the smallest size. After optimization of the pegRNAs and incorporation with nick sgRNAs, the mini-PE delivered up to 10% precise editing at target sites in human and mouse cells. It also edited the mouse Hsf1 gene in the mouse retina precisely after delivery with adeno-associated viruses (AAVs), although the editing efficiency was lower than 1%. We will focus on improving the editing efficiency of mini-PE and exploiting its therapeutic potential against human genetic diseases.© 2023 The Author(s).