High throughput single-cell detection of multiplex CRISPR-edited gene modifications

作者：Elisa ten Hacken, Kendell Clement, Shuqiang Li, María Hernández‐Sánchez, Robert Redd, Shu Wang, David Ruff, Michaela Gruber, Kaitlyn Baranowski, Jose Jacob, James M. Flynn, Keith Jones, Donna Neuberg, Kenneth J. Livak, Luca Pinello, Catherine J. Wu · 发表于：Genome biology · 年份：2020 · DOI：10.1186/s13059-020-02174-1 · 被引用次数：34 · 研究领域：CRISPR and Genetic Engineering、Single-cell and spatial transcriptomics、CAR-T cell therapy research

CRISPR-Cas9 gene editing has transformed our ability to rapidly interrogate the functional impact of somatic mutations in human cancers. Droplet-based technology enables the analysis of Cas9-introduced gene edits in thousands of single cells. Using this technology, we analyze Ba/F3 cells engineered to express single or multiplexed loss-of-function mutations recurrent in chronic lymphocytic leukemia. Our approach reliably quantifies mutational co-occurrences, zygosity status, and the occurrence of Cas9 edits at single-cell resolution.