Long-read individual-molecule sequencing reveals CRISPR-induced genetic heterogeneity in human ESCs
Long-read individual-molecule sequencing reveals CRISPR-induced genetic heterogeneity in human ESCs
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DOI：
10.1186/s13059-020-02143-8
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发表时间：
2020-08-24
期刊：
影响因子：
12.3
通讯作者：
Li, Mo
中科院分区：
文献类型：
--
作者：
Bi, Chongwei;Wang, Lin;Li, Mo
量化细胞群体的遗传异质性对于理解生物系统至关重要。我们开发了一种通用的方法来标记单个DNA分子，用于对不同类型的罕见变异进行单碱基分辨率单体型分辨定量表征，频率低至4 x 10（-5），使用短读或长读测序平台。它提供了第一个定量证据，证明在人类胚胎干细胞中CRISPR-Cas9诱导的双链断裂修复后，持续的非随机大结构变异和靶位点单核苷酸变异的增加。
Quantifying the genetic heterogeneity of a cell population is essential to understanding of biological systems. We develop a universal method to label individual DNA molecules for single-base-resolution haplotype-resolved quantitative characterization of diverse types of rare variants, with frequency as low as 4 x 10(-5), using both short- or long-read sequencing platforms. It provides the first quantitative evidence of persistent nonrandom large structural variants and an increase in single-nucleotide variants at the on-target locus following repair of double-strand breaks induced by CRISPR-Cas9 in human embryonic stem cells.