期刊
NATURE COMMUNICATIONS
卷 8, 期 -, 页码 -出版社
NATURE PORTFOLIO
DOI: 10.1038/s41467-017-01836-2
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资金
- Programa de Movilidad en Areas de Investigacion priorizadas por la Consejeria de Igualdad, Salud y Politicas Sociales de la Junta de Andalucia
- NIH [R21 HD073768, R01 HD074078, GM103789, GM102251, GM101108, GM081602]
- Swiss National Science Foundation [P2GEP3_148600, R01 HD081379, 4R33HL120783]
- National Institute Of General Medical Sciences of the National Institutes of Health [R35GM122580]
- Edward Mallinckrodt Jr Foundation
- HHMI Faculty Scholars program
- March of Dimes
- Yale Scholars Program
- Marine Biological Laboratory in Woods Hole
- Australian National Health and Medical Research Council [APP1090875]
- Swiss National Science Foundation (SNF) [P2GEP3_148600] Funding Source: Swiss National Science Foundation (SNF)
Cpf1 is a novel class of CRISPR-Cas DNA endonucleases, with a wide range of activity across different eukaryotic systems. Yet, the underlying determinants of this variability are poorly understood. Here, we demonstrate that LbCpf1, but not AsCpf1, ribonucleoprotein complexes allow efficient mutagenesis in zebrafish and Xenopus. We show that temperature modulates Cpf1 activity by controlling its ability to access genomic DNA. This effect is stronger on AsCpf1, explaining its lower efficiency in ectothermic organisms. We capitalize on this property to show that temporal control of the temperature allows post-translational modulation of Cpf1-mediated genome editing. Finally, we determine that LbCpf1 significantly increases homology-directed repair in zebrafish, improving current approaches for targeted DNA integration in the genome. Together, we provide a molecular understanding of Cpf1 activity in vivo and establish Cpf1 as an efficient and inducible genome engineering tool across ectothermic species.
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