| RRC ID |
90085
|
| Author |
Wu Y, Wang J, Zhang Z, Shang M, Wang S, Li Y, Li G, Lu S, Ji K, Wang X, Zhang X.
|
| Title |
Triple base editor catalyzes saturation mutation of adenine, cytidine, and guanine.
|
| Journal |
Nucleic Acids Res
|
| Abstract |
Current base editors act on a maximum of two base substrates and generate limited base conversions or transversions, hindering their applicability for inducing DNA sequence diversity. Here, we engineered a triple base editor (named ACG-BEs) using a fusion of adenine base editor with high A/C catalytic activity and evolved N-methylpurine DNA glycosylase. ACG-BEs enables efficient, multiplexed saturation mutagenesis across adenine (A), cytosine (C), and guanine (G), achieving conversion efficiencies of up to 80.5% for A-to-G/C/T, 75.8% for C-to-T/G/A, and 63.4% for G-to-C/T/A in HEK293T cells. Leveraging ACG-BEs, we identify novel mutations in the HBG1/2 promoter region that confer efficient activation of γ-globin expression in HUDEP-2 cells-a promising advancement for therapeutic strategies targeting hemoglobinopathies. These findings highlight ACG-BEs as a cutting-edge platform for multiplexed saturation mutagenesis, offering broad applications in genetic screening and therapeutic base mutation introduction through enhanced DNA sequence diversity.
|
| Volume |
54(2)
|
| Published |
2026-1-14
|
| DOI |
10.1093/nar/gkaf1423
|
| PII |
8425338
|
| PMID |
41533583
|
| PMC |
PMC12802905
|
| MeSH |
Adenine* / chemistry
Adenine* / metabolism
Cytidine* / chemistry
Cytidine* / metabolism
DNA Glycosylases* / genetics
DNA Glycosylases* / metabolism
Guanine* / chemistry
Guanine* / metabolism
HEK293 Cells
Humans
Mutagenesis*
Mutation*
|
| Resource |
| Human and Animal Cells |
HUDEP-2(RCB4557) |
