title: A High-Throughput Strategy for Dissecting Mammalian Genetic Interactions.
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.
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, New York, United States of America.
authors.forename: Victoria B
authors.initials: VB
authors.lastname: Stockman
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, New York, United States of America.|Department of Microbiology and Immunology, Columbia University Medical Center, New York, New York, United States of America.
authors.forename: Lila
authors.initials: L
authors.lastname: Ghamsari
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, New York, United States of America.|Department of Biochemistry, Columbia University Medical Center, New York, New York, United States of America.
authors.forename: Gorka
authors.initials: G
authors.lastname: Lasso
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, New York, United States of America.|Department of Biochemistry, Columbia University Medical Center, New York, New York, United States of America.
authors.forename: Barry
authors.initials: B
authors.lastname: Honig
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, New York, United States of America.|Department of Microbiology and Immunology, Columbia University Medical Center, New York, New York, United States of America.
authors.forename: Sagi D
authors.initials: SD
authors.lastname: Shapira
authors.affiliation: Department of Systems Biology, Columbia University Medical Center, New York, New York, United States of America.|Department of Pathology and Cell Biology, Columbia University Medical Center, New York, New York, United States of America.
authors.forename: Harris H
authors.identifier: http://orcid.org/0000-0003-2164-4318
authors.initials: HH
authors.lastname: Wang
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D000072669:Gene Editing; D053263:Gene Regulatory Networks; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D011401:Promoter Regions, Genetic; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems