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Multiplexed pooled library screening with Cpf1

By Jintan Liu, Sanjana Srinivasan, Chieh-Yuan Li, I-Lin Ho, Gang Wang, Wantong Yao, Angela Kay Deem, Chris Bristow, Traver Hart, Giulio Draetta

Posted 19 Nov 2018
bioRxiv DOI: 10.1101/473959 (published DOI: 10.1038/s41467-019-10963-x)

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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