Details
| Original language | English |
|---|---|
| Article number | 18 |
| Journal | Genome biology |
| Volume | 2025 |
| Issue number | 26 |
| Publication status | Published - 3 Feb 2025 |
Abstract
Base editors are precise editing tools that employ deaminases to modify target DNA bases. The DYW-family of cytosine deaminases is structurally and phylogenetically distinct and might be harnessed for genome editing tools. We report a novel CRISPR/Cas9-cytosine base editor using SsdA, a DYW-like deaminase and bacterial toxin. A G103S mutation in SsdA enhances C-to-T editing efficiency while reducing its toxicity. Truncations result in an extraordinarily small enzyme. The SsdA-base editor efficiently converts C-to-T in rice and barley protoplasts and induces mutations in rice plants and mammalian cells. The engineered SsdA is a highly efficient genome editing tool.
Keywords
- Gene Editing, Cytosine/metabolism, CRISPR-Cas Systems, Oryza/genetics, Humans, Bacterial Toxins/genetics, Cytosine Deaminase/genetics, Animals, Hordeum/genetics, Protoplasts/metabolism, HEK293 Cells, Mutation, Base editing, Crop improvement, Gene therapy, CRISPR
ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Ecology, Evolution, Behavior and Systematics
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Genome biology, Vol. 2025, No. 26, 18, 03.02.2025.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Engineering a bacterial toxin deaminase from the DYW-family into a novel cytosine base editor for plants and mammalian cells
AU - Zhang, Dingbo
AU - Parth, Fiona
AU - Matos da Silva, Laura
AU - Ha, Teng-Cheong
AU - Schambach, Axel
AU - Boch, Jens
N1 - © 2025. The Author(s).
PY - 2025/2/3
Y1 - 2025/2/3
N2 - Base editors are precise editing tools that employ deaminases to modify target DNA bases. The DYW-family of cytosine deaminases is structurally and phylogenetically distinct and might be harnessed for genome editing tools. We report a novel CRISPR/Cas9-cytosine base editor using SsdA, a DYW-like deaminase and bacterial toxin. A G103S mutation in SsdA enhances C-to-T editing efficiency while reducing its toxicity. Truncations result in an extraordinarily small enzyme. The SsdA-base editor efficiently converts C-to-T in rice and barley protoplasts and induces mutations in rice plants and mammalian cells. The engineered SsdA is a highly efficient genome editing tool.
AB - Base editors are precise editing tools that employ deaminases to modify target DNA bases. The DYW-family of cytosine deaminases is structurally and phylogenetically distinct and might be harnessed for genome editing tools. We report a novel CRISPR/Cas9-cytosine base editor using SsdA, a DYW-like deaminase and bacterial toxin. A G103S mutation in SsdA enhances C-to-T editing efficiency while reducing its toxicity. Truncations result in an extraordinarily small enzyme. The SsdA-base editor efficiently converts C-to-T in rice and barley protoplasts and induces mutations in rice plants and mammalian cells. The engineered SsdA is a highly efficient genome editing tool.
KW - Gene Editing
KW - Cytosine/metabolism
KW - CRISPR-Cas Systems
KW - Oryza/genetics
KW - Humans
KW - Bacterial Toxins/genetics
KW - Cytosine Deaminase/genetics
KW - Animals
KW - Hordeum/genetics
KW - Protoplasts/metabolism
KW - HEK293 Cells
KW - Mutation
KW - Base editing
KW - Crop improvement
KW - Gene therapy
KW - CRISPR
UR - http://www.scopus.com/inward/record.url?scp=85217570512&partnerID=8YFLogxK
U2 - 10.1186/s13059-025-03478-w
DO - 10.1186/s13059-025-03478-w
M3 - Article
C2 - 39901278
VL - 2025
JO - Genome biology
JF - Genome biology
SN - 1474-760X
IS - 26
M1 - 18
ER -