June
(2022-04-30 14:29):
#paper https://doi.org/10.1038/s41392-022-00936-w该研究通过将 RT 与核酸酶野生型 Cas9 (WT-PE) 融合来进行大规模基因组操作,设计了一种新的先导编辑系统。与传统的先导编辑器(PE2)不同,这种新系统同时在目标位点引入了一个 DSB 和一个 3' 延伸的瓣,然后通过内源机制将它们整合到基因组中。当它与配对的 pegRNA 结合时,WT-PE 实现了高效的大规模基因组编辑,包括大片段缺失和染色体易位。因此, WT-PE 系统可能有助于建模或治疗与大片段畸变相关的疾病。
IF:40.800Q1
Signal transduction and targeted therapy,
2022-04-20.
DOI: 10.1038/s41392-022-00936-w
PMID: 35440051
PMCID:PMC9018734
WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
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Abstract:
Large scale genomic aberrations including duplication, deletion, translocation, and other structural changes are the cause of a subtype of hereditary genetic disorders and contribute to onset or progress of cancer. The current prime editor, PE2, consisting of Cas9-nickase and reverse transcriptase enables efficient editing of genomic deletion and insertion, however, at small scale. Here, we designed a novel prime editor by fusing reverse transcriptase (RT) to nuclease wild-type Cas9 (WT-PE) to edit large genomic fragment. WT-PE system simultaneously introduced a double strand break (DSB) and a single 3' extended flap in the target site. Coupled with paired prime editing guide RNAs (pegRNAs) that have complementary sequences in their 3' terminus while target different genomic regions, WT-PE produced bi-directional prime editing, which enabled efficient and versatile large-scale genome editing, including large fragment deletion up to 16.8 megabase (Mb) pairs and chromosomal translocation. Therefore, our WT-PE system has great potential to model or treat diseases related to large-fragment aberrations.
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