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Pooled library screening with multiplexed Cpf1 library.

ABSTRACT: Capitalizing on the inherent multiplexing capability of AsCpf1, we developed a multiplexed, high-throughput screening strategy that minimizes library size without sacrificing gene targeting efficiency. We demonstrated that AsCpf1 can be used for functional genomics screenings and that an AsCpf1-based multiplexed library performs similarly as compared to currently available monocistronic CRISPR/Cas9 libraries, with only one vector required for each gene. We construct the smallest whole-genome CRISPR knock-out library, Mini-human, for the human genome (n?=?17,032 constructs targeting 16,977 protein-coding genes), which performs favorably compared to conventional Cas9 libraries.

SUBMITTER: Liu J

PROVIDER: S-EPMC6637147 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Pooled library screening with multiplexed Cpf1 library.

Liu Jintan J Srinivasan Sanjana S Li Chieh-Yuan CY Ho I-Lin IL Rose Johnathon J Shaheen MennatAllah M Wang Gang G Yao Wantong W Deem Angela A Bristow Chris C Hart Traver T Draetta Giulio G

Nature communications 20190717 1

Capitalizing on the inherent multiplexing capability of AsCpf1, we developed a multiplexed, high-throughput screening strategy that minimizes library size without sacrificing gene targeting efficiency. We demonstrated that AsCpf1 can be used for functional genomics screenings and that an AsCpf1-based multiplexed library performs similarly as compared to currently available monocistronic CRISPR/Cas9 libraries, with only one vector required for each gene. We construct the smallest whole-genome CRI ...[more]

PMID: 31316073

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