CAMERS-B: CRISPR/Cpf1 assisted multiple-genes editing and regulation system for Bacillus subtilis.

Biotechnology and Bioengineering
Yaokang WuYang Gu

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.

References

Mar 19, 2003·Proceedings of the National Academy of Sciences of the United States of America·Shunji NakanoPeter Zuber
Aug 2, 2005·Journal of Molecular Biology·María I Martínez-JiménezSilvia Ayora
Nov 22, 2011·Journal of Biotechnology·Trang Thi Phuong PhanWolfgang Schumann
Feb 1, 2015·Applied and Environmental Microbiology·Yu JiangSheng Yang
May 5, 2017·Nature Communications·Yu JiangSheng Yang
Jul 12, 2017·Frontiers in Microbiology·Younju SoSoo-Keun Choi
Jun 29, 2018·Nature Communications·Chen DongJesse G Zalatan
Oct 1, 2018·World Journal of Microbiology & Biotechnology·Kun-Qiang HongZhi-Wen Wang
Oct 6, 2018·Nucleic Acids Research·Shuyi Zhang, Christopher A Voigt

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Citations

Feb 23, 2021·Frontiers in Cell and Developmental Biology·Baisong TongDawei Zhang
Mar 24, 2021·Applied Microbiology and Biotechnology·Meliawati MeliawatiJochen Schmid
Aug 7, 2021·Frontiers in Bioengineering and Biotechnology·Chaoyong LiaoBingkun Zhang
Aug 15, 2021·Nucleic Acids Research·Wenliang HaoZhemin Zhou
Jan 12, 2021·Briefings in Bioinformatics·Hui PengJinyan Li

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