Evolution of CRISPR-associated endonucleases as inferred from resurrected proteins.
Journal
Nature microbiology
ISSN: 2058-5276
Titre abrégé: Nat Microbiol
Pays: England
ID NLM: 101674869
Informations de publication
Date de publication:
01 2023
01 2023
Historique:
received:
24
05
2022
accepted:
07
10
2022
pubmed:
3
1
2023
medline:
10
1
2023
entrez:
2
1
2023
Statut:
ppublish
Résumé
Clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9 is an effector protein that targets invading DNA and plays a major role in the prokaryotic adaptive immune system. Although Streptococcus pyogenes CRISPR-Cas9 has been widely studied and repurposed for applications including genome editing, its origin and evolution are poorly understood. Here, we investigate the evolution of Cas9 from resurrected ancient nucleases (anCas) in extinct firmicutes species that last lived 2.6 billion years before the present. We demonstrate that these ancient forms were much more flexible in their guide RNA and protospacer-adjacent motif requirements compared with modern-day Cas9 enzymes. Furthermore, anCas portrays a gradual palaeoenzymatic adaptation from nickase to double-strand break activity, exhibits high levels of activity with both single-stranded DNA and single-stranded RNA targets and is capable of editing activity in human cells. Prediction and characterization of anCas with a resurrected protein approach uncovers an evolutionary trajectory leading to functionally flexible ancient enzymes.
Identifiants
pubmed: 36593295
doi: 10.1038/s41564-022-01265-y
pii: 10.1038/s41564-022-01265-y
pmc: PMC10324499
mid: NIHMS1902005
doi:
Substances chimiques
CRISPR-Associated Protein 9
EC 3.1.-
Endonucleases
EC 3.1.-
RNA, Guide, CRISPR-Cas Systems
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
77-90Subventions
Organisme : NIMHD NIH HHS
ID : L60 MD003721
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL142494
Pays : United States
Informations de copyright
© 2023. The Author(s), under exclusive licence to Springer Nature Limited.
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