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
Dec 6, 2008·BMC Biotechnology·Huanting Liu, James H Naismith
Nov 22, 2011·Journal of Biotechnology·Trang Thi Phuong PhanWolfgang Schumann
Feb 25, 2014·Metabolic Engineering·Yanfeng LiuJian Chen
Oct 14, 2014·Cell·Marvin E TanenbaumRonald D Vale
Feb 1, 2015·Applied and Environmental Microbiology·Yu JiangSheng Yang
Jul 15, 2015·PloS One·Satohiko MurayamaTaku Oshima
Oct 1, 2015·Cell·Bernd ZetscheFeng Zhang
May 5, 2017·Nature Communications·Yu JiangSheng Yang
Jul 12, 2017·Frontiers in Microbiology·Younju SoSoo-Keun Choi
Dec 9, 2017·Bioresource Technology·Wenlong MaLong Liu
May 19, 2018·Metabolic Engineering·Yang GuLong Liu
Jun 27, 2018·ACS Synthetic Biology·Dingyu LiuXueming Zhao
Jun 29, 2018·Nature Communications·Chen DongJesse G Zalatan
Aug 24, 2018·ACS Synthetic Biology·Tengfei NiuLong Liu
Sep 4, 2018·Metabolic Engineering·Yaokang WuLong Liu
Oct 1, 2018·World Journal of Microbiology & Biotechnology·Kun-Qiang HongZhi-Wen Wang
Oct 6, 2018·Nucleic Acids Research·Shuyi Zhang, Christopher A Voigt
Dec 7, 2018·Synthetic and Systems Biotechnology·Chensi MiaoChunbo Lou
Jan 8, 2019·Bioinformatics·Houxiang Zhu, Chun Liang
Jan 31, 2019·Nucleic Acids Research·Lei FuAnding Zhang
Feb 2, 2019·ACS Synthetic Biology·Maike KortmannMichael Bott
Feb 16, 2019·Nucleic Acids Research·Zhenghui LuGuimin Zhang
Oct 2, 2019·Biotechnology for Biofuels·Dingyu LiuXueming Zhao
Dec 5, 2019·Nucleic Acids Research·Yaokang WuLong Liu
Citations
Sep 29, 2020·Frontiers in Bioengineering and Biotechnology·Wenliang HaoZhemin Zhou
Sep 5, 2020·Microbial Cell Factories·Yuan SuDawei Zhang
Dec 8, 2020·Frontiers in Microbiology·Bin YeXin Yan
Jan 13, 2021·Bioresource Technology·Han YangJian Chen
Feb 23, 2021·Frontiers in Cell and Developmental Biology·Baisong TongDawei Zhang
Mar 24, 2021·Applied Microbiology and Biotechnology·Meliawati MeliawatiJochen Schmid
May 7, 2021·Bioresource Technology·Jiajia YouZhiming Rao
May 12, 2021·Biotechnology Advances·Seung-Woon JungSeung Min Yoo
Aug 7, 2021·Frontiers in Bioengineering and Biotechnology·Chaoyong LiaoBingkun Zhang
Aug 15, 2021·Nucleic Acids Research·Wenliang HaoZhemin Zhou
Aug 17, 2020·ACS Synthetic Biology·Yaokang WuLong Liu
Jan 6, 2021·ACS Synthetic Biology·Zhijie QinJingwen Zhou
Sep 2, 2021·Bioresource Technology·Xinzhu MaoLong Liu
Jan 12, 2021·Briefings in Bioinformatics·Hui PengJinyan Li
Sep 7, 2021·Frontiers in Microbiology·Aihua DengTingyi Wen