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Implementing CRISPR-Cas12a for Efficient Genome Editing in Yarrowia lipolytica
1Department of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore County, Baltimore, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|April 13, 2021
Summary
We present a new CRISPR-Cas12a genome editing protocol for the yeast Yarrowia lipolytica. This method expands genetic engineering tools for producing valuable chemicals, complementing existing CRISPR-Cas9 systems.
Area of Science:
- Biotechnology
- Synthetic Biology
- Microbial Engineering
Background:
- Yarrowia lipolytica is a key oleaginous yeast for industrial chemical production.
- Effective metabolic engineering requires robust genetic tools.
- Current tools include homologous recombination, random integration, episomal plasmids, and CRISPR-Cas9.
Purpose of the Study:
- To describe a protocol for multiplexed genome editing in Yarrowia lipolytica using CRISPR-Cas12a.
- To establish CRISPR-Cas12a as an efficient genome-editing tool for Y. lipolytica.
Main Methods:
- Utilized the CRISPR effector Cas12a for genome engineering.
- Delivered AsCas12a and crRNA expression via a single plasmid.
- Implemented multiplexed genome editing strategies.
Main Results:
- Demonstrated the feasibility of CRISPR-Cas12a for genome editing in Y. lipolytica.
- Showcased Cas12a's unique features like T-rich PAM and self-processing crRNA.
Conclusions:
- CRISPR-Cas12a expands the genetic toolbox for Y. lipolytica.
- This system is complementary to CRISPR-Cas9, offering new possibilities for metabolic engineering.
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