PEM-seq检测基因组稳定性数据集

本数据集描述了一种新方法，即引物延伸介导的测序 (PEM-seq)，它可以灵敏地检测 CRISPR/Cas9 脱靶位点，并通过量化在靶位点的 DNA 异常事件来评估其编辑效率。PEM-seq数据证明，我们设计了一种高保真 Cas9 变体 FeCas9，它具有与野生型相似的靶向能力，但脱靶活性极低。此外，我们证明了xCas9确实具有更广泛的PAM序列识别序列。我们还发现 AcrIIA4 抑制剂可以抑制 SpCas9 的靶向和脱靶活性，但它抑制 SpCas9 对脱靶位点的切割不如在靶向位点有效。最后，我们认为 PEM-seq 适用于优化基因组编辑策略以用于临床目的或构建动物模型。

This dataset describes a novel method, primer extension-mediated sequencing (PEM-seq), which enables sensitive detection of CRISPR/Cas9 off-target sites and evaluation of editing efficiency by quantifying DNA aberrant events at on-target sites. PEM-seq data demonstrate that we engineered a high-fidelity Cas9 variant, FeCas9, which exhibits on-target activity comparable to that of wild-type Cas9 but with extremely low off-target activity. Furthermore, we verified that xCas9 indeed recognizes a broader spectrum of PAM sequences. We also discovered that the AcrIIA4 inhibitor can suppress both the on-target and off-target activities of SpCas9, yet it inhibits SpCas9-mediated cleavage at off-target sites less efficiently than at on-target sites. Finally, we conclude that PEM-seq is suitable for optimizing genome editing strategies for clinical applications or animal model construction.

搜集汇总

背景与挑战

背景概述

该数据集基于PEM-seq方法，专注于检测CRISPR/Cas9基因编辑中的脱靶位点和评估编辑效率，关键发现包括高保真Cas9变体FeCas9的低脱靶活性、xCas9的广泛PAM识别能力以及AcrIIA4抑制剂对SpCas9活性的影响。数据集由北京大学发布，数据量2.5GB，包含19个文件，旨在支持基因组编辑策略优化和临床或动物模型应用。