Tags

Type your tag names separated by a space and hit enter

CAMERS-B: CRISPR/Cpf1 assisted multiple-genes editing and regulation system for Bacillus subtilis.
Biotechnol Bioeng. 2020 Jun; 117(6):1817-1825.BB

Abstract

The clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) systems have been widely used in genome editing and transcriptional regulation. In this study, by engineering the Francisella novicida U112 CRISPR/Cpf1 system, a powerful tool called CRISPR/Cpf1 assisted multiple-genes editing and regulation system for B. subtilis was constructed for engineering Bacillus subtilis, and a synthetic oligos mediated assembly of CRISPR RNA (crRNA) array method was created to build crRNA array. This system can achieve the double genes in-frame knocking out, multiple point mutations (up to six), or single gene insertion at a time with 100% efficiency. In addition, transcriptional regulation systems were also developed using the DNase deactivated Cas protein (dCpf1) and a transcription factor RemA, which can implement repression and activation on multiple-genes concurrently. Finally, as a proof-of-concept demonstration, the synthesis pathways of N-acetylglucosamine and acetoin in B. subtilis were engineered by using this system. Overall, we provide effective tools for genome editing and metabolic engineering of B. subtilis cell factories to produce various biochemicals.

Authors+Show Affiliations

Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32129468

Citation

Wu, Yaokang, et al. "CAMERS-B: CRISPR/Cpf1 Assisted Multiple-genes Editing and Regulation System for Bacillus Subtilis." Biotechnology and Bioengineering, vol. 117, no. 6, 2020, pp. 1817-1825.
Wu Y, Liu Y, Lv X, et al. CAMERS-B: CRISPR/Cpf1 assisted multiple-genes editing and regulation system for Bacillus subtilis. Biotechnol Bioeng. 2020;117(6):1817-1825.
Wu, Y., Liu, Y., Lv, X., Li, J., Du, G., & Liu, L. (2020). CAMERS-B: CRISPR/Cpf1 assisted multiple-genes editing and regulation system for Bacillus subtilis. Biotechnology and Bioengineering, 117(6), 1817-1825. https://doi.org/10.1002/bit.27322
Wu Y, et al. CAMERS-B: CRISPR/Cpf1 Assisted Multiple-genes Editing and Regulation System for Bacillus Subtilis. Biotechnol Bioeng. 2020;117(6):1817-1825. PubMed PMID: 32129468.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - CAMERS-B: CRISPR/Cpf1 assisted multiple-genes editing and regulation system for Bacillus subtilis. AU - Wu,Yaokang, AU - Liu,Yanfeng, AU - Lv,Xueqin, AU - Li,Jianghua, AU - Du,Guocheng, AU - Liu,Long, Y1 - 2020/03/16/ PY - 2019/10/19/received PY - 2020/02/20/revised PY - 2020/03/02/accepted PY - 2020/3/5/pubmed PY - 2020/3/5/medline PY - 2020/3/5/entrez KW - Bacillus subtilis KW - CRISPR/Cpf1 KW - CRISPRa KW - CRISPRi KW - genome editing KW - synthetic biology SP - 1817 EP - 1825 JF - Biotechnology and bioengineering JO - Biotechnol. Bioeng. VL - 117 IS - 6 N2 - The clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) systems have been widely used in genome editing and transcriptional regulation. In this study, by engineering the Francisella novicida U112 CRISPR/Cpf1 system, a powerful tool called CRISPR/Cpf1 assisted multiple-genes editing and regulation system for B. subtilis was constructed for engineering Bacillus subtilis, and a synthetic oligos mediated assembly of CRISPR RNA (crRNA) array method was created to build crRNA array. This system can achieve the double genes in-frame knocking out, multiple point mutations (up to six), or single gene insertion at a time with 100% efficiency. In addition, transcriptional regulation systems were also developed using the DNase deactivated Cas protein (dCpf1) and a transcription factor RemA, which can implement repression and activation on multiple-genes concurrently. Finally, as a proof-of-concept demonstration, the synthesis pathways of N-acetylglucosamine and acetoin in B. subtilis were engineered by using this system. Overall, we provide effective tools for genome editing and metabolic engineering of B. subtilis cell factories to produce various biochemicals. SN - 1097-0290 UR - https://www.unboundmedicine.com/medline/citation/32129468/CAMERS-B:_CRISPR/Cpf1_assisted_multiple-genes_editing_and_regulation_system_for_Bacillus_subtilis L2 - https://doi.org/10.1002/bit.27322 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.