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A High-Throughput Strategy for Dissecting Mammalian Genetic Interactions

Stockman, Victoria B.; Ghamsari, Lila; Lasso Cabrera, Gorka; Honig, Barry; Shapira, Sagi; Wang, Harris H.

Comprehensive delineation of complex cellular networks requires high-throughput interrogation of genetic interactions. To address this challenge, we describe the development of a multiplex combinatorial strategy to assess pairwise genetic interactions using CRISPR-Cas9 genome editing and next-generation sequencing. We characterize the performance of combinatorial genome editing and analysis using different promoter and gRNA designs and identified regions of the chimeric RNA that are compatible with next-generation sequencing preparation and quantification. This approach is an important step towards elucidating genetic networks relevant to human diseases and the development of more efficient Cas9-based therapeutics.

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Academic Units
Systems Biology
Biochemistry and Molecular Biophysics
Microbiology and Immunology
Pathology and Cell Biology
Publisher
Public Library of Science
Published Here
February 8, 2017
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