Glycosylase base editors enable C-to-A and C-to-G base changes
| Autor: | Marcus A. Price, Ju Li, Susan J. Rosser, Muzi Hu, Siwei Li, Changhao Bi, Dongdong Zhao, Xiuqing Xin, Xueli Zhang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Chemistry DNA repair APOBEC1 Biomedical Engineering Bioengineering Cytidine Cytidine deaminase medicine.disease_cause Applied Microbiology and Biotechnology Molecular biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine DNA glycosylase Uracil-DNA glycosylase medicine Molecular Medicine AP site Escherichia coli 030217 neurology & neurosurgery 030304 developmental biology Biotechnology |
| Zdroj: | Zhao, D, Li, J, Li, S, Xin, X, Hu, M, Price, M A, Rosser, S J, Bi, C & Zhang, X 2020, ' Glycosylase base editors enable C-to-A and C-to-G base changes ', Nature Biotechnology, vol. 39, pp. 35-40 . https://doi.org/10.1038/s41587-020-0592-2 |
| DOI: | 10.1038/s41587-020-0592-2 |
| Popis: | Current base editors (BEs) catalyze only base transitions (C to T and A to G) and cannot produce base transversions. Here we present BEs that cause C-to-A transversions in Escherichia coli and C-to-G transversions in mammalian cells. These glycosylase base editors (GBEs) consist of a Cas9 nickase, a cytidine deaminase and a uracil-DNA glycosylase (Ung). Ung excises the U base created by the deaminase, forming an apurinic/apyrimidinic (AP) site that initiates the DNA repair process. In E. coli, we used activation-induced cytidine deaminase (AID) to construct AID-nCas9-Ung and found that it converts C to A with an average editing specificity of 93.8% ± 4.8% and editing efficiency of 87.2% ± 6.9%. For use in mammalian cells, we replaced AID with rat APOBEC1 (APOBEC-nCas9-Ung). We tested APOBEC-nCas9-Ung at 30 endogenous sites, and we observed C-to-G conversions with a high editing specificity at the sixth position of the protospacer between 29.7% and 92.2% and an editing efficiency between 5.3% and 53.0%. APOBEC-nCas9-Ung supplements the current adenine and cytidine BEs (ABE and CBE, respectively) and could be used to target G/C disease-causing mutations. New base editors change C to A in bacteria and C to G in mammalian cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|