Liang Ronghong, Wang Shan, Cai Yibo, Li Zhenyu, Li Ka Ming, Wei Jingjing, Sun Chao, Zhu Haocheng, Chen Kunling, Gao Caixia
New Cornerstone Science Laboratory, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing, China.
Nat Commun. 2025 May 31;16(1):5057. doi: 10.1038/s41467-025-59120-7.
Prime editors are restricted to performing precise edits downstream of cleavage sites, thereby limiting their editing scope. Therefore, we develop inverse prime editors (iPEs) that act upstream of the nickase cleavage site by replacing nCas9-H840A with nCas9-D10A, but the editing efficiencies are limited. To address this limitation, we develop circular RNA-mediated iPEs (ciPEs), achieving editing efficiencies ranging from 0.1% to 24.7%. Further optimization using Rep-X helicase increases editing efficiencies to a range of 2.7%-55.4%. The Rep-X-assisted ciPE system thus expands the scope of editing and improves efficiencies at genomic sites that are previously difficult to target. The Rep-X-assisted ciPE system will complement canonical PE system in enabling more extensive and efficient editing across a wider range of the human genome.
引导编辑器仅限于在切割位点下游进行精确编辑，从而限制了它们的编辑范围。因此，我们开发了反向引导编辑器（iPE），通过用nCas9-D10A替换nCas9-H840A，使其在切口酶切割位点上游起作用，但编辑效率有限。为了解决这一限制，我们开发了环状RNA介导的iPE（ciPE），实现了0.1%至24.7%的编辑效率。使用Rep-X解旋酶进行进一步优化可将编辑效率提高到2.7%-55.4%的范围。因此，Rep-X辅助的ciPE系统扩大了编辑范围，并提高了以前难以靶向的基因组位点的编辑效率。Rep-X辅助的ciPE系统将补充传统的引导编辑器系统，以便在更广泛的人类基因组中实现更广泛、更高效的编辑。
