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Long-read Individual-molecule Sequencing Reveals CRISPR-induced Genetic Heterogeneity in Human ESCs

By Chongwei Bi, Lin Wang, Baolei Yuan, Xuan Zhou, Yu Pang, Yu Li, Sheng Wang, Yuhong Xin Gao, Yanyi Huang, Mo Li

Posted 11 Feb 2020
bioRxiv DOI: 10.1101/2020.02.10.942151

Accurately 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 analyzing diverse types of rare genetic variants, with frequency as low as 4x10-5, using short- or long-read sequencing. It enables base-resolution haplotype-resolved quantitative characterization of rare variants. It provides the first quantitative evidence of persistent nonrandom large deletions and insertions following DNA repair of double-strand breaks induced by CRISPR-Cas9 in human pluripotent stem cells.

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