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:10.1371/journal.pone.0167617

A high-throughput strategy for dissecting mammalian genetic interactions

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

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

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 CRISPRCas9 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.

Original language	English (US)
Article number	e0167617
Journal	PloS one
Volume	11
Issue number	12
DOIs	https://doi.org/10.1371/journal.pone.0167617
State	Published - Dec 2016
Externally published	Yes

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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 CRISPRCas9 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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AU - Ghamsari, Lila

AU - Lasso, Gorka

AU - Honig, Barry

AU - Shapira, Sagi D.

AU - Wang, Harris H.

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N2 - 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 CRISPRCas9 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.

AB - 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 CRISPRCas9 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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A high-throughput strategy for dissecting mammalian genetic interactions

Abstract

ASJC Scopus subject areas

UN SDGs

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