Programmable DNA pyrimidine base editing via engineered uracil-DNA glycosylase.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
30 Jul 2024
30 Jul 2024
Historique:
received:
19
03
2024
accepted:
27
06
2024
medline:
31
7
2024
pubmed:
31
7
2024
entrez:
30
7
2024
Statut:
epublish
Résumé
DNA base editing technologies predominantly utilize engineered deaminases, limiting their ability to edit thymine and guanine directly. In this study, we successfully achieve base editing of both cytidine and thymine by leveraging the translesion DNA synthesis pathway through the engineering of uracil-DNA glycosylase (UNG). Employing structure-based rational design, exploration of homologous proteins, and mutation screening, we identify a Deinococcus radiodurans UNG mutant capable of effectively editing thymine. When fused with the nickase Cas9, the engineered DrUNG protein facilitates efficient thymine base editing at endogenous sites, achieving editing efficiencies up to 55% without enrichment and exhibiting minimal cellular toxicity. This thymine base editor (TBE) exhibits high editing specificity and significantly restores IDUA enzyme activity in cells derived from patients with Hurler syndrome. TBEs represent efficient, specific, and low-toxicity approaches to base editing with potential applications in treating relevant diseases.
Identifiants
pubmed: 39080265
doi: 10.1038/s41467-024-50012-w
pii: 10.1038/s41467-024-50012-w
doi:
Substances chimiques
Uracil-DNA Glycosidase
EC 3.2.2.-
DNA
9007-49-2
Thymine
QR26YLT7LT
CRISPR-Associated Protein 9
EC 3.1.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
6397Subventions
Organisme : National Natural Science Foundation of China (National Science Foundation of China)
ID : NSFC31930016
Informations de copyright
© 2024. The Author(s).
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