Automated identification of sequence-tailored Cas9 proteins using massive metagenomic data.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
29 10 2022
29 10 2022
Historique:
received:
31
08
2022
accepted:
17
10
2022
pubmed:
31
10
2022
medline:
2
11
2022
entrez:
30
10
2022
Statut:
epublish
Résumé
The identification of the protospacer adjacent motif (PAM) sequences of Cas9 nucleases is crucial for their exploitation in genome editing. Here we develop a computational pipeline that was used to interrogate a massively expanded dataset of metagenome and virome assemblies for accurate and comprehensive PAM predictions. This procedure allows the identification and isolation of sequence-tailored Cas9 nucleases by using the target sequence as bait. As proof of concept, starting from the disease-causing mutation P23H in the RHO gene, we find, isolate and experimentally validate a Cas9 which uses the mutated sequence as PAM. Our PAM prediction pipeline will be instrumental to generate a Cas9 nuclease repertoire responding to any PAM requirement.
Identifiants
pubmed: 36309502
doi: 10.1038/s41467-022-34213-9
pii: 10.1038/s41467-022-34213-9
pmc: PMC9617884
doi:
Substances chimiques
CRISPR-Associated Protein 9
EC 3.1.-
RNA, Guide
0
Endonucleases
EC 3.1.-
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
6474Subventions
Organisme : NCI NIH HHS
ID : U01 CA230551
Pays : United States
Informations de copyright
© 2022. The Author(s).
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