A High-Throughput Strategy for Dissecting Mammalian Genetic Interactions.

Stockman, Victoria B; Ghamsari, Lila; Lasso, Gorka; Honig, Barry; Shapira, Sagi D; Wang, Harris H · PLoS One · 2016

basic_science · Level V

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Abstract

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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