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

Genome Biology

Kendell Clement¹  Luca Pinello¹  Shuqiang Li²  Donna Neuberg³  Robert Redd³  Kaitlyn Baranowski⁴  Michaela Gruber⁵  Elisa ten Hacken⁶  Catherine J. Wu⁷  María Hernández-Sánchez⁸  Keith W. Jones⁹  Shu Wang⁹  David Ruff⁹  James Flynn⁹  Jose Jacob⁹  Kenneth J. Livak^1,10 

[1] Broad Institute of MIT and Harvard, Cambridge, MA, USA;Molecular Pathology Unit, Center for Cancer Research and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, MA, USA;Department of Pathology, Harvard Medical School, Boston, MA, USA;Broad Institute of MIT and Harvard, Cambridge, MA, USA;Translational Immunogenomics Lab, Dana-Farber Cancer Institute, Boston, MA, USA;Department of Data Science, Dana-Farber Cancer Institute, Boston, MA, USA;Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA;Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA;Department of Internal Medicine I, Division of Hematology and Hemostaseology, Medical University of Vienna, Vienna, Austria;Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA;Harvard Medical School, Boston, MA, USA;Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA;Harvard Medical School, Boston, MA, USA;Broad Institute of MIT and Harvard, Cambridge, MA, USA;Department of Medicine, Brigham and Women’s Hospital, Boston, MA, USA;Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA;IBSAL, IBMCC-Cancer Research Center, University of Salamanca, Salamanca, Spain;Mission Bio, Incorporated, South San Francisco, CA, USA;Translational Immunogenomics Lab, Dana-Farber Cancer Institute, Boston, MA, USA;

关键词: Genetics;    Single cell;    Genome editing;    Loss-of-function;    Mutation;    CRISPR-Cas9;    chronic lymphocytic leukemia;   

DOI  :  10.1186/s13059-020-02174-1

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY