CRISPR-dCas9 Mediated Cytosine Deaminase Base Editing in Bacillus subtilis
| Autor: | Yang Liu, Sili Yu, Meng Wang, Marcus A. Price, Changhao Bi, Xiaomeng Ni, Yanmei Guo, Susan J. Rosser, Yu Wang |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Biomedical Engineering Bacillus subtilis 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) 03 medical and health sciences chemistry.chemical_compound bacillus subtilis Genome editing 010608 biotechnology Activation-induced (cytidine) deaminase genome editing CRISPR cytidine deaminase 030304 developmental biology Genetics 0303 health sciences biology Cas9 Cytosine deaminase General Medicine Cytidine deaminase biology.organism_classification chemistry biology.protein CRISPR/dCas9 DNA |
| Zdroj: | Yu, S, Price, M A, Wang, Y, Liu, Y, Guo, Y, Ni, X, Rosser, S J, Bi, C & Wang, M 2020, ' CRISPR-dCas9 mediated cytosine deaminase base editing in bacillus subtilis ', ACS Synthetic Biology, vol. 9, no. 7, pp. 1781-1789 . https://doi.org/10.1021/acssynbio.0c00151 |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.0c00151 |
| Popis: | Base editing technology based on clustered regularly interspaced short palindromic repeats/associated protein 9 (CRISPR/Cas9) is a recent addition to the family of CRISPR technologies. Compared with the traditional CRISPR/Cas9 technology, it does not rely on DNA double strand break and homologous recombination, and can realize gene inactivation and point mutation more quickly and simply. Herein, we first developed a base editing method for genome editing in Bacillus subtilis utilizing CRISPR/dCas9 (a fully nuclease-deficient mutant of Cas9 from S. pyogenes) and activation-induced cytidine deaminase (AID). This method achieved three and four loci simultaneous editing with editing efficiency up to 100% and 50%, respectively. Our base editing system in B. subtilis has a 5 nt editing window, which is similar to previously reported base editing in other microorganisms. We demonstrated that the plasmid curing rate is almost 100%, which is advantageous for multiple rounds of genome engineering in B. subtilis. Finally, we applied multiplex genome editing to generate a B. subtilis 168 mutant strain with eight inactive extracellular protease genes in just two rounds of base editing and plasmid curing, suggesting that it is a powerful tool for gene manipulation in B. subtilis and industrial applications in the future. |
| Databáze: | OpenAIRE |
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