High throughput single-cell detection of multiplex CRISPR-edited gene modifications
Elisa ten Hacken, Kendell Clement, Shuqiang Li, Maria Hernandez-Sanchez, Robert Redd, Shu Wang, David Ruff, Michaela Gruber, Kaitlyn Baranowski, Jose Jacob, James Flynn, Keith W. Jones, Donna Neuberg, Kenneth J. Livak, Luca Pinello, Catherine J. Wu
Genome Biology
Oct 2020
Abstract
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.
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Area
Heme
Institution Type
Academia
Indication / Modality
Chronic Lymphocytic Leukemia (CLL)
Goal of Study
Genome Editing
Key Genes
TP53, CHD2, BIRC3, MGA, SAMHD1
PAD Project
No
Analytes Assessed
InDels, SNV
Sample Storage
Fresh Frozen
Sample Prep
Whole Cells
Sample Type
Cell Line
Tissue / Organ
Cell Line
Species
Human
Panel Used
Custom
Proof Point Demonstrated
Better than Bulk, Zygosity, Co-occurrence
