Development of deaminase-free T-to-S base editor and C-to-G base editor by engineered human uracil DNA glycosylase.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
08 Jun 2024
08 Jun 2024
Historique:
received:
18
03
2024
accepted:
31
05
2024
medline:
9
6
2024
pubmed:
9
6
2024
entrez:
8
6
2024
Statut:
epublish
Résumé
DNA base editors enable direct editing of adenine (A), cytosine (C), or guanine (G), but there is no base editor for direct thymine (T) editing currently. Here we develop two deaminase-free glycosylase-based base editors for direct T editing (gTBE) and C editing (gCBE) by fusing Cas9 nickase (nCas9) with engineered human uracil DNA glycosylase (UNG) variants. By several rounds of structure-informed rational mutagenesis on UNG in cultured human cells, we obtain gTBE and gCBE with high activity of T-to-S (i.e., T-to-C or T-to-G) and C-to-G conversions, respectively. Furthermore, we conduct parallel comparison of gTBE/gCBE with those recently developed using other protein engineering strategies, and find gTBE/gCBE show the outperformance. Thus, we provide several base editors, gTBEs and gCBEs, with corresponding engineered UNG variants, broadening the targeting scope of base editors.
Identifiants
pubmed: 38851742
doi: 10.1038/s41467-024-49343-5
pii: 10.1038/s41467-024-49343-5
doi:
Substances chimiques
Uracil-DNA Glycosidase
EC 3.2.2.-
CRISPR-Associated Protein 9
EC 3.1.-
Cytosine
8J337D1HZY
Thymine
QR26YLT7LT
Guanine
5Z93L87A1R
DNA
9007-49-2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4897Informations de copyright
© 2024. The Author(s).
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