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Plant Gene Knockout and Knockdown by CRISPR-Cpf1 (Cas12a) Systems
Yingxiao Zhang1, Yong Zhang2, Yiping Qi3,4
1Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
Summary
A new CRISPR-Cpf1 (Cas12a) vector assembly system enables rapid genome editing in plants. This efficient tool achieved high editing efficiency and gene expression reduction in rice and Arabidopsis.
Area of Science:
- Molecular Biology
- Biotechnology
- Plant Science
Background:
- CRISPR-Cpf1 (Cas12a) is a versatile class II type V endonuclease utilized for genome editing across various biological systems.
- Efficient and user-friendly CRISPR-Cpf1 vector assembly is crucial for advancing plant genome engineering.
Purpose of the Study:
- To develop a fast, efficient, and user-friendly system for assembling CRISPR-Cpf1 expression vectors.
- To enable precise crRNA processing and customizable component modification for diverse plant species and experimental objectives.
Main Methods:
- Utilized a Pol II promoter to drive co-expression of Cpf1 and crRNA.
- Incorporated hammerhead (HH) and hepatitis delta virus (HDV) ribozyme RNAs flanking the crRNA for accurate processing.
- Demonstrated system adaptability for different plant species and research aims.
Main Results:
- Achieved nearly 100% editing efficiency in rice.
- Obtained up to 90% gene expression decrease in Arabidopsis.
- Validated the system's effectiveness and ease of use in plant genome editing.
Conclusions:
- The developed CRISPR-Cpf1 vector assembly system offers a rapid and efficient method for plant genome editing.
- The system's modular design and precise crRNA processing contribute to high editing efficiencies and gene silencing.
- This tool facilitates advancements in plant biotechnology and genetic research.
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