小柯机器人

美国马萨诸塞大学医学院Scot A. Wolfe等研究人员，合作使用PE-tag对体外和体内的先导编辑非靶标位点进行全基因组分析。2023年5月8日，《自然—方法学》杂志在线发表了这项成果。

研究人员描述了一种全基因组的方法，用于识别潜在的主要编辑脱靶点，被称为PE-tag。这种方法依赖于在先导编辑活性的位点上附着或插入一个扩增标签，进而让它们被识别。PE-tag能够在体外使用提取的基因组DNA，在哺乳动物细胞系和成年小鼠肝脏中进行全基因组的脱靶点分析。PE-tag成分可以以各种形式传递，用于检测脱靶位点。这项研究与先前描述的先导编辑系统的高特异性一致，但研究人员发现脱靶编辑率受先导编辑向导RNA设计的影响。PE-tag代表了一种可获得的、快速和敏感的方法，可用于全基因组鉴定先导编辑活性和评估先导编辑安全性。

据介绍，先导编辑具有广泛的潜在研究和临床应用。然而，划定其全基因组编辑活性的方法通常依赖于间接的全基因组编辑评估，或近似识别序列的计算预测。

附：英文原文

Title: Genome-wide profiling of prime editor off-target sites in vitro and in vivo using PE-tag

Author: Liang, Shun-Qing, Liu, Pengpeng, Ponnienselvan, Karthikeyan, Suresh, Sneha, Chen, Zexiang, Kramme, Christian, Chatterjee, Pranam, Zhu, Lihua Julie, Sontheimer, Erik J., Xue, Wen, Wolfe, Scot A.

Issue&Volume: 2023-05-08

Abstract: Prime editors have a broad range of potential research and clinical applications. However, methods to delineate their genome-wide editing activities have generally relied on indirect genome-wide editing assessments or the computational prediction of near-cognate sequences. Here we describe a genome-wide approach for the identification of potential prime editor off-target sites, which we call PE-tag. This method relies on the attachment or insertion of an amplification tag at sites of prime editor activity to allow their identification. PE-tag enables genome-wide profiling of off-target sites in vitro using extracted genomic DNA, in mammalian cell lines and in the adult mouse liver. PE-tag components can be delivered in a variety of formats for off-target site detection. Our studies are consistent with the high specificity previously described for prime editor systems, but we find that off-target editing rates are influenced by prime editing guide RNA design. PE-tag represents an accessible, rapid and sensitive approach for the genome-wide identification of prime editor activity and the evaluation of prime editor safety.

DOI: 10.1038/s41592-023-01859-2

Source: https://www.nature.com/articles/s41592-023-01859-2

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex