Adaptation and application of a two-plasmid inducible CRISPR-Cas9 system in Clostridium beijerinckii

Methods : a Companion to Methods in Enzymology
Mamou DialloA M López-Contreras

Abstract

Recent developments in CRISPR technologies have opened new possibilities for improving genome editing tools dedicated to the Clostridium genus. In this study we adapted a two-plasmid tool based on this technology to enable scarless modification of the genome of two reference strains of Clostridium beijerinckii producing an Acetone/Butanol/Ethanol (ABE) or an Isopropanol/Butanol/Ethanol (IBE) mix of solvents. In the NCIMB 8052 ABE-producing strain, inactivation of the SpoIIE sporulation factor encoding gene resulted in sporulation-deficient mutants, and this phenotype was reverted by complementing the mutant strain with a functional spoIIE gene. Furthermore, the fungal cellulase-encoding celA gene was inserted into the C. beijerinckii NCIMB 8052 chromosome, resulting in mutants with endoglucanase activity. A similar two-plasmid approach was next used to edit the genome of the natural IBE-producing strain C. beijerinckii DSM 6423, which has never been genetically engineered before. Firstly, the catB gene conferring thiamphenicol resistance was deleted to make this strain compatible with our dual-plasmid editing system. As a proof of concept, our dual-plasmid system was then used in C. beijerinckii DSM 6423 ΔcatB to remove the end...Continue Reading

References

Dec 1, 1986·Microbiological Reviews·D T Jones, D R Woods
Nov 1, 1985·Applied and Environmental Microbiology·M HermannJ P Vandecasteele
Feb 11, 2003·Trends in Biochemical Sciences·Thomas E WilsonPhillip L Palmbos
Jul 27, 2005·Journal of Industrial Microbiology & Biotechnology·M González-PajueloI Vasconcelos
Jun 28, 2006·Proceedings of the National Academy of Sciences of the United States of America·Hildur V ColotJay C Dunlap
May 1, 2007·Applied and Environmental Microbiology·Florence MingardonHenri-Pierre Fierobe
Jul 31, 2007·Journal of Microbiological Methods·John T HeapNigel P Minton
Mar 4, 2008·Applied and Environmental Microbiology·Stephen A RodriguezKarl E Klose
Apr 2, 2008·Annals of the New York Academy of Sciences·Peter Dürre
Oct 27, 2009·Applied and Environmental Microbiology·Burkhard OtteStefan Jennewein
Nov 7, 2009·Journal of Microbiological Methods·John T HeapNigel P Minton
Nov 26, 2010·Applied and Environmental Microbiology·Andrea H HartmanStephen B Melville
Feb 22, 2011·Applied and Environmental Microbiology·Marco A J SiemerinkServé W M Kengen
May 17, 2011·Applied Microbiology and Biotechnology·Shrikant A SurvaseTom Granström
Oct 4, 2011·Applied and Environmental Microbiology·Hirofumi NariyaAkinobu Okabe
Jan 18, 2012·Applied Microbiology and Biotechnology·Wouter KuitGerrit Eggink
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jan 30, 2015·Microbiology and Molecular Biology Reviews : MMBR·Mohab A Al-HinaiEleftherios T Papoutsakis
Feb 15, 2015·Journal of Biotechnology·Yi WangHans P Blaschek
Apr 26, 2015·Applied and Environmental Microbiology·Tao XuJizhong Zhou
Feb 8, 2016·Applied Microbiology and Biotechnology·H Máté de GérandoA M López-Contreras
Oct 26, 2016·Biotechnology for Biofuels·Shilpa NagarajuSéan Dennis Simpson
Mar 14, 2017·Biotechnology for Biofuels·Anja PoehleinPeter Dürre
Jun 15, 2017·Journal of Microbiological Methods·François WaselsNicolas Lopes Ferreira
Feb 8, 2018·Current Opinion in Biotechnology·Ioannis MougiakosRichard van Kranenburg
Apr 11, 2018·BMC Genomics·Hadrien Máté de GérandoNicolas Lopes Ferreira

❮ Previous
Next ❯

Citations

Oct 13, 2020·Frontiers in Microbiology·Mamou DialloAna M López-Contreras
Apr 27, 2021·Applied Microbiology and Biotechnology·Mamou DialloAna M López-Contreras
May 1, 2021·International Journal of Molecular Sciences·Tom Wilding-SteelePhilippe Soucaille
Oct 7, 2021·Nucleic Acids Research·Constantinos PatiniosRaymond H J Staals

❮ Previous
Next ❯

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.