PLoS ONE (Jan 2016)

A High-Throughput Strategy for Dissecting Mammalian Genetic Interactions.

  • Victoria B Stockman,
  • Lila Ghamsari,
  • Gorka Lasso,
  • Barry Honig,
  • Sagi D Shapira,
  • Harris H Wang

DOI
https://doi.org/10.1371/journal.pone.0167617
Journal volume & issue
Vol. 11, no. 12
p. e0167617

Abstract

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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.