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Genome Editing of Corynebacterium glutamicum Using CRISPR-Cpf1 System
Zhiqiang Wen1, Fenghui Qian2, Jiao Zhang3,4
1School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, China.
Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2022
Summary
Efficient genome editing in Corynebacterium glutamicum was achieved using Francisella novicida (Fn) CRISPR-Cpf1 systems. This breakthrough enables rapid, iterative genetic modifications for industrial applications.
Area of Science:
- Microbiology
- Synthetic Biology
- Biotechnology
Background:
- Corynebacterium glutamicum is a key microbial host with significant industrial potential.
- Efficient genetic modification tools are lacking for C. glutamicum, hindering its application.
- The Streptococcus pyogenes (Sp) CRISPR-Cas9 system is ineffective in C. glutamicum.
Purpose of the Study:
- To develop and describe efficient genome editing tools for Corynebacterium glutamicum.
- To enable rapid and iterative genetic modifications in C. glutamicum.
Main Methods:
- Development of two Francisella novicida (Fn) CRISPR-Cpf1 assisted systems.
- Application of these systems for genome editing via homologous recombination in C. glutamicum.
Main Results:
- Demonstrated successful genome editing in C. glutamicum using Fn CRISPR-Cpf1 systems.
- Achieved N iterative rounds of genome editing in 3N + 4 or 3N + 2 days, showcasing high efficiency.
Conclusions:
- The developed Fn CRISPR-Cpf1 systems are effective for genome editing in C. glutamicum.
- These tools significantly accelerate genetic modification processes, unlocking C. glutamicum's industrial potential.
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