Programmable RNA-guided DNA endonucleases are widespread in eukaryotes and their viruses
Open accessAbstract
Programmable RNA-guided DNA nucleases perform numerous roles in prokaryotes, but the extent of their spread outside prokaryotes is unclear. Fanzors, the eukaryotic homolog of prokaryotic TnpB proteins, have been detected in genomes of eukaryotes and large viruses, but their activity and functions in eukaryotes remain unknown. Here, we characterize Fanzors as RNA-programmable DNA endonucleases, using biochemical and cellular evidence. We found diverse Fanzors that frequently associate with various eukaryotic transposases. Reconstruction of Fanzors evolution revealed multiple radiations of RuvC-containing TnpB homologs in eukaryotes. Fanzor genes captured introns and proteins acquired nuclear localization signals, indicating extensive, long-term adaptation to functioning in eukaryotic cells. Fanzor nucleases contain a rearranged catalytic site of the RuvC domain, similar to a distinct subset of TnpBs, and lack collateral cleavage activity. We demonstrate that Fanzors can be harnessed for genome editing in human cells, highlighting the potential of these widespread eukaryotic RNA-guided nucleases for biotechnology applications.
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“Fanzors are homologous to the prokaryotic TnpB proteins and have been detected in various eukaryotic genomes, suggesting a widespread presence beyond prokaryotic systems (Jiang et al., 2023).”
Influential
- Authors
- Jiang, K., Lim, J., Sgrizzi, S., Trinh, M., Kayabolen, A., Yutin, N., Bao, W., Kato, K., Koonin, E. V., Gootenberg, J. S., & Abudayyeh, O. O.
- Affiliations
- McGovern Institute for Brain Research (US)
- McGovern Institute for Brain Research at MIT Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
- National Institutes of Health (US)
- Genetic Information Research Institute (US)
- Tokyo Medical and Dental University (JP)
- Corresponding author
- Jiang, Kaiyi
- Lim, Justin
- Gootenberg, Jonathan S.
- Abudayyeh, Omar O.
- ORCID
- Venue
- Science Advances
- Published
- 2023-09-29
- Volume
- 9
- Issue
- 39
- Pages
- eadk0171-eadk0171
- ISSN
- 2375-2548 (ISSN-L)
- PMC
- PMC10530073
- OpenAlex
- W4387092069
- Cited by
- 60
- References
- 34
- Citations by year
- 2026: 16 · 2025: 20 · 2024: 20 · 2023: 4
- FWCI
- 5.32
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by-nc
- Locations
- Science Advances (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (1r56hg011857-01)
- National Institute on Aging (R01AG074932)
- National Human Genome Research Institute (R56-HG011857)
- National Institute of Allergy and Infectious Diseases (R21AI149694)
- National Institute of Biomedical Imaging and Bioengineering (R01EB031957)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- RNA-guided DNA endonuclease, eukaryotic viruses, genome editing, nuclear localization signal, collateral cleavage, human cells
- MeSH terms
- Deoxyribonuclease I, Deoxyribonucleases, Humans, RNA, Viruses, Eukaryota
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