Peking University Yichengqi Research Group Nature published an article: Double-pegRNA-mediated high-efficiency guided editing

On October 28, 2021, the research group of Professor Yi Chengqi from Peking University School of Life Sciences and Peking University-Tsinghua Life Sciences Joint Center published a study titled " Increasing the efficiency and precision of prime editing with guide RNA pairs " in Nature Chemical Biology .
The paper developed a new strategy for efficient guided editing in human-derived cells based on double pegRNA .

Prime Editing can realize all 12 kinds of base substitutions, and can also realize the insertion and deletion of small fragments
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Guided editing is based on the CRISPR/Cas9 system, fusing nCas9 (H840A) with reverse transcriptase to obtain a new fusion protein

In this study, two pegRNAs targeted to the double-stranded DNA were designed to edit the same site in the genome, so as to improve the efficiency of guided editing
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Since the two pegRNAs contain homologous 3'ends, this strategy is named "HOPE" (HOmologous 3'Extensions Mediated Prime Editor)

Diagram of the working principle of the HOPE strategy

In summary, this study uses the dual pegRNA strategy to significantly improve the efficiency of guided editing
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Through verification on a series of endogenous sites and different cell lines, it is confirmed that the HOPE strategy provides a new choice for guided editing with a high balance of efficiency and accuracy

Yi Chengqi is the corresponding author of this article
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Doctoral student Zhuang Yuan from the School of Life Sciences, doctoral student Liu Jiangle from the Peking University-Tsinghua Life Sciences Joint Center, and doctoral student Wu Hao are the co-first authors of this article