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Systematically investigating the key features of the nuclease deactivated Cpf1 for tunable multiplex genetic regulation

Chensi Miao, Huiwei Zhao, Long Qian, Chunbo Lou
doi: https://doi.org/10.1101/297903
Chensi Miao
Institute of Microbiology, CAS;
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Huiwei Zhao
Institute of Microbiology, CAS;
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Long Qian
Department of Biology and Center for Genomics and Systems Biology, New York University;
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Chunbo Lou
Chinese Academy of Science
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  • For correspondence: louchunbo@im.ac.cn
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Abstract

With a unique crRNA processing capability, the CRISPR associated Cpf1 protein holds great potential for multiplex gene regulation. Unlike the well-studied Cas9 protein, however, conversion of Cpf1 to a transcription regulator and its related properties have not been systematically explored yet. In this study, we investigated the mutation schemes and crRNA requirements for the nuclease deactivated Cpf1 (dCpf1). By shortening the direct repeat sequence, we obtained genetically stable crRNA co-transcripts and improved gene repression with multiplex targeting. A screen of diversity-enriched PAM library was designed to investigate the PAM-dependency of gene regulation by dCpf1 from Francisella novicida and Lachnospiraceae bacterium. We found novel PAM patterns that elicited strong or medium gene repressions. Using a computational algorithm, we predicted regulatory outputs for all possible PAM sequences, which spanned a large dynamic range that could be leveraged for regulatory purposes. These newly identified features will facilitate the efficient design of CRISPR-dCpf1 based systems for tunable multiplex gene regulation.

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  • Posted April 9, 2018.

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Systematically investigating the key features of the nuclease deactivated Cpf1 for tunable multiplex genetic regulation
Chensi Miao, Huiwei Zhao, Long Qian, Chunbo Lou
bioRxiv 297903; doi: https://doi.org/10.1101/297903
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Systematically investigating the key features of the nuclease deactivated Cpf1 for tunable multiplex genetic regulation
Chensi Miao, Huiwei Zhao, Long Qian, Chunbo Lou
bioRxiv 297903; doi: https://doi.org/10.1101/297903

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