Pooled library screening with multiplexed Cpf1 library

Jintan Liu (), Sanjana Srinivasan, Chieh-Yuan Li, I-Lin Ho, Johnathon Rose, MennatAllah Shaheen, Gang Wang, Wantong Yao, Angela Deem, Chris Bristow, Traver Hart and Giulio Draetta ()
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Jintan Liu: The University of Texas MD Anderson Cancer Center
Sanjana Srinivasan: The University of Texas MD Anderson Cancer Center
Chieh-Yuan Li: The University of Texas MD Anderson Cancer Center
I-Lin Ho: The University of Texas MD Anderson Cancer Center
Johnathon Rose: The University of Texas MD Anderson Cancer Center
MennatAllah Shaheen: The University of Texas MD Anderson Cancer Center
Gang Wang: The University of Texas MD Anderson Cancer Center
Wantong Yao: The University of Texas MD Anderson Cancer Center
Angela Deem: The University of Texas MD Anderson Cancer Center
Chris Bristow: The University of Texas MD Anderson Cancer Center
Traver Hart: The University of Texas MD Anderson Cancer Center
Giulio Draetta: The University of Texas MD Anderson Cancer Center

Nature Communications, 2019, vol. 10, issue 1, 1-10

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-019-10963-x

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