Multiplexed pooled library screening with Cpf1

Jintan Liu; Sanjana Srinivasan; Chieh-Yuan Li; I-Lin Ho; Johnathon Lynn Rose; MennatAllah Shaheen; Gang Wang; Wantong Yao; Angela K. Deem; Chris Bristow; Traver Hart; Giulio Francesco Draetta

doi:10.1038/s41467-019-10963-x

Systems BiologyNature CommunicationsJul 17, 2019

Peer-reviewed journal article

Multiplexed pooled library screening with Cpf1

Jintan Liu iD, Sanjana Srinivasan iD, Chieh-Yuan Li iD, I-Lin Ho iD, Johnathon Lynn Rose iD, MennatAllah Shaheen iD, Gang Wang iD, Wantong Yao iD, Angela K. Deem iD, Chris Bristow, Traver Hart iD, Giulio Francesco Draetta iD

Abstract

RNA interference and CRISPR/Cas9-based pooled library screens have revolutionized the field of functional genomics. However, currently available pooled library screens face a trade-off between library effectiveness and library complexity. We developed a multiplexed, high-throughput screening strategy based on an optimized AsCpf1 nuclease that minimizes library size without sacrificing gene targeting efficiency. Our AsCpf1-based multiplexed library performed similarly well compared to currently available CRISPR/Cas9 libraries, but with a single polycistronic crRNA clone targeting each gene. With this strategy, we constructed the smallest whole-genome knock-out library available, \"Mini-human\" for the human genome, which is one-fourth the size of the smallest CRISPR library currently available.

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