Development of TALE-adenine base editors in plants
- authored by
- Dingbo Zhang, Jens Boch
- Abstract
Base editors enable precise nucleotide changes at targeted genomic loci without requiring double-stranded DNA breaks or repair templates. TALE-adenine base editors (TALE-ABEs) are genome editing tools, composed of a DNA-binding domain from transcription activator-like effectors (TALEs), an engineered adenosine deaminase (TadA8e), and a cytosine deaminase domain (DddA), that allow A•T-to-G•C editing in human mitochondrial DNA. However, the editing ability of TALE-ABEs in plants apart from chloroplast DNA has not been described, so far, and the functional role how DddA enhances TadA8e is still unclear. We tested a series of TALE-ABEs with different deaminase fusion architectures in Nicotiana benthamiana and rice. The results indicate that the double-stranded DNA-specific cytosine deaminase DddA can boost the activities of single-stranded DNA-specific deaminases (TadA8e or APOBEC3A) on double-stranded DNA. We analysed A•T-to-G•C editing efficiencies in a β-glucuronidase reporter system and showed precise adenine editing in genomic regions with high product purity in rice protoplasts. Furthermore, we have successfully regenerated rice plants with A•T-to-G•C mutations in the chloroplast genome using TALE-ABE. Consequently, TALE-adenine base editors provide alternatives for crop improvement and gene therapy by editing nuclear or organellar genomes.
- Organisation(s)
-
Institute of Plant Genetics
- Type
- Article
- Journal
- Plant Biotechnology Journal
- Volume
- 22
- Pages
- 1067-1077
- No. of pages
- 11
- ISSN
- 1467-7644
- Publication date
- 17.04.2024
- Publication status
- Published
- Peer reviewed
- Yes
- ASJC Scopus subject areas
- Agronomy and Crop Science, Biotechnology, Plant Science
- Electronic version(s)
-
https://doi.org/10.1111/pbi.14246 (Access:
Open)