Tags

Type your tag names separated by a space and hit enter

CRISPR-dCas9 Mediated Cytosine Deaminase Base Editing in Bacillus subtilis.
ACS Synth Biol. 2020 Jul 17; 9(7):1781-1789.AS

Abstract

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.

Authors+Show Affiliations

College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.Centre for Synthetic and Systems Biology and UK Centre for Mammalian Synthetic Biology, School of Biological Sciences, University of Edinburgh, Edinburgh EH8 9YL, U.K.College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.Centre for Synthetic and Systems Biology and UK Centre for Mammalian Synthetic Biology, School of Biological Sciences, University of Edinburgh, Edinburgh EH8 9YL, U.K.College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32551562

Citation

Yu, Sili, et al. "CRISPR-dCas9 Mediated Cytosine Deaminase Base Editing in Bacillus Subtilis." ACS Synthetic Biology, vol. 9, no. 7, 2020, pp. 1781-1789.
Yu S, Price MA, Wang Y, et al. CRISPR-dCas9 Mediated Cytosine Deaminase Base Editing in Bacillus subtilis. ACS Synth Biol. 2020;9(7):1781-1789.
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, 9(7), 1781-1789. https://doi.org/10.1021/acssynbio.0c00151
Yu S, et al. CRISPR-dCas9 Mediated Cytosine Deaminase Base Editing in Bacillus Subtilis. ACS Synth Biol. 2020 Jul 17;9(7):1781-1789. PubMed PMID: 32551562.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - CRISPR-dCas9 Mediated Cytosine Deaminase Base Editing in Bacillus subtilis. AU - Yu,Sili, AU - Price,Marcus A, AU - Wang,Yu, AU - Liu,Yang, AU - Guo,Yanmei, AU - Ni,Xiaomeng, AU - Rosser,Susan J, AU - Bi,Changhao, AU - Wang,Meng, Y1 - 2020/07/01/ PY - 2020/6/20/pubmed PY - 2020/6/20/medline PY - 2020/6/20/entrez KW - Bacillus subtilis KW - CRISPR/dCas9 KW - cytidine deaminase KW - genome editing SP - 1781 EP - 1789 JF - ACS synthetic biology JO - ACS Synth Biol VL - 9 IS - 7 N2 - 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. SN - 2161-5063 UR - https://www.unboundmedicine.com/medline/citation/32551562/CRISPR-dCas9_Mediated_Cytosine_Deaminase_Base_Editing_in_Bacillus_subtilis L2 - https://doi.org/10.1021/acssynbio.0c00151 DB - PRIME DP - Unbound Medicine ER -
Try the Free App:
Prime PubMed app for iOS iPhone iPad
Prime PubMed app for Android
Prime PubMed is provided
free to individuals by:
Unbound Medicine.