Pooled library screening with multiplexed Cpf1 library

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-base...

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Bibliographic Details
Published in:	Nature communications 2019-07, Vol.10 (1), p.3144-10, Article 3144
Main Authors:	Liu, Jintan, Srinivasan, Sanjana, Li, Chieh-Yuan, Ho, I-Lin, Rose, Johnathon, Shaheen, MennatAllah, Wang, Gang, Yao, Wantong, Deem, Angela, Bristow, Chris, Hart, Traver, Draetta, Giulio
Format:	Article
Language:	eng
Subjects:	Clustered Regularly Interspaced Short Palindromic Repeats - genetics CRISPR CRISPR-Associated Protein 9 - genetics CRISPR-Cas Systems - genetics Gene Editing Gene Library Gene targeting Genomes Genomics High-throughput screening Humans Libraries Multiplexing RNA, Guide, CRISPR-Cas Systems - chemistry Screening
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Summary:	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.
ISSN:	2041-1723 2041-1723