The University of Calgary, Cumming School of Medicine, Department of Biochemistry and Molecular Biology, 3330 Hospital Drive N.W., Calgary, AB T2N 1N4, Canada; Robson DNA Science Centre, Calgary, AB, Canada; Arnie Charbonneau Cancer Institute, Calgary, AB, Canada.
The University of Calgary, Cumming School of Medicine, Department of Biochemistry and Molecular Biology, 3330 Hospital Drive N.W., Calgary, AB T2N 1N4, Canada; Robson DNA Science Centre, Calgary, AB, Canada; Arnie Charbonneau Cancer Institute, Calgary, AB, Canada.
STAR Protoc. 2024 Sep 20;5(3):103307. doi: 10.1016/j.xpro.2024.103307. Epub 2024 Sep 16.
Prime editing is a highly versatile CRISPR-based genome editing technology that allows for the precise installation of desired genetic variants. This protocol describes how to use One-pot DTECT to assess prime editing efficiency in human cells. Key steps include conducting prime editing, extracting genomic DNA, performing AcuI-tagging PCR, capturing genetic signatures, and detecting captured signatures through qualitative, quantitative, and visual methods. One-pot DTECT enables same-day detection of targeted genetic signatures introduced by precision genome editing technologies using off-the-shelf reagents. For complete details on the use and execution of this protocol, please refer to Baudrier et al..
碱基编辑是一种基于 CRISPR 的高度通用的基因组编辑技术，可实现精确地安装所需的遗传变体。本方案描述了如何使用 One-pot DTECT 来评估人细胞中的碱基编辑效率。主要步骤包括进行碱基编辑、提取基因组 DNA、进行 AcuI 标记 PCR、捕获遗传特征以及通过定性、定量和可视化方法检测捕获的特征。One-pot DTECT 使用即用型试剂实现了当天检测使用精确基因组编辑技术引入的靶向遗传特征。如需详细了解本方案的使用和执行情况，请参考 Baudrier 等人的研究。
