CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing
Abstract
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.
- Authors:
-
- Yale Univ. School of Medicine, New Haven, CT (United States)
- Univ. of California, Berkeley, CA (United States)
- Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1560545
- Grant/Contract Number:
- AC02-05CH11231; R21 HD073768; R01 HD074078; GM103789; GM102251; GM101108; GM081602
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nature Communications
- Additional Journal Information:
- Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2041-1723
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Moreno-Mateos, Miguel A., Fernandez, Juan P., Rouet, Romain, Vejnar, Charles E., Lane, Maura A., Mis, Emily, Khokha, Mustafa K., Doudna, Jennifer A., and Giraldez, Antonio J. CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. United States: N. p., 2017.
Web. doi:10.1038/s41467-017-01836-2.
Moreno-Mateos, Miguel A., Fernandez, Juan P., Rouet, Romain, Vejnar, Charles E., Lane, Maura A., Mis, Emily, Khokha, Mustafa K., Doudna, Jennifer A., & Giraldez, Antonio J. CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. United States. doi:10.1038/s41467-017-01836-2.
Moreno-Mateos, Miguel A., Fernandez, Juan P., Rouet, Romain, Vejnar, Charles E., Lane, Maura A., Mis, Emily, Khokha, Mustafa K., Doudna, Jennifer A., and Giraldez, Antonio J. Fri .
"CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing". United States. doi:10.1038/s41467-017-01836-2. https://www.osti.gov/servlets/purl/1560545.
@article{osti_1560545,
title = {CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing},
author = {Moreno-Mateos, Miguel A. and Fernandez, Juan P. and Rouet, Romain and Vejnar, Charles E. and Lane, Maura A. and Mis, Emily and Khokha, Mustafa K. and Doudna, Jennifer A. and Giraldez, Antonio J.},
abstractNote = {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.},
doi = {10.1038/s41467-017-01836-2},
journal = {Nature Communications},
number = 1,
volume = 8,
place = {United States},
year = {2017},
month = {12}
}
Web of Science
Figures / Tables:

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