PEM-seq comprehensively quantifies DNA repair outcomes during gene-editing and DSB repair

STAR Protocols(2022)

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摘要
The repair products of double-stranded DNA breaks (DSBs) are crucial for investigating the mechanism underlying DNA damage repair as well as evaluating the safety and efficiency of gene-editing; however, a comprehensively quantitative assay remains to be established. Here, we describe the step-by-step instructions of the primer extension-mediated sequencing (PEM-seq), followed by the framework of data processing and statistical analysis. PEM-seq presents a full spectrum of repair outcomes for both genome-editing-induced and endogenous DSBs in mouse and human cells.
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关键词
Bioinformatics,CRISPR,High Throughput Screening,Molecular Biology,Sequence analysis,Sequencing
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