Long-read individual-molecule sequencing reveals CRISPR-induced genetic heterogeneity in human ESCs
Chongwei Bi, Lin Wang, Baolei Yuan, Xuan Zhou, Yu Li, Sheng Wang, Yuhong Pang, Xin Gao, Yanyi Huang, Mo Li
Abstract
Abstract 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 × 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.
Topics & Concepts
BiologyCRISPRGeneticsHuman geneticsLocus (genetics)Computational biologyPopulationDNASingle molecule real time sequencingHaplotypeEmbryonic stem cellGenetic heterogeneityDNA sequencingGenePhenotypeGenotypeDNA sequencerSociologyDemographyCRISPR and Genetic EngineeringSingle-cell and spatial transcriptomicsAdvanced biosensing and bioanalysis techniques