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Volume 12
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10.3390/plants12071478
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Review

Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

by
Junming Zhou
1,
Xinchao Luan
1,
Yixuan Liu
1,
Lixue Wang
1,
Jiaxin Wang
1,
Songnan Yang
2,
Shuying Liu
1,
Jun Zhang
2,
Huijing Liu
1,* and
Dan Yao
1,*
1
College of Life Sciences, Jilin Agricultural University, Changchun 130118, China
2
College of Agronomy, Jilin Agricultural University, Changchun 130118, China
*
Authors to whom correspondence should be addressed.
Plants 2023, 12(7), 1478; https://doi.org/10.3390/plants12071478
Submission received: 22 February 2023 / Revised: 21 March 2023 / Accepted: 22 March 2023 / Published: 28 March 2023
(This article belongs to the Special Issue Plant Chromosome Biology and Genomics for Breeding)
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Abstract

Following recent developments and refinement, CRISPR-Cas9 gene-editing technology has become increasingly mature and is being widely used for crop improvement. The application of CRISPR/Cas9 enables the generation of transgene-free genome-edited plants in a short period and has the advantages of simplicity, high efficiency, high specificity, and low production costs, which greatly facilitate the study of gene functions. In plant molecular breeding, the gene-editing efficiency of the CRISPR-Cas9 system has proven to be a key step in influencing the effectiveness of molecular breeding, with improvements in gene-editing efficiency recently becoming a focus of reported scientific research. This review details strategies and methods for improving the efficiency of CRISPR/Cas9 gene editing in plant molecular breeding, including Cas9 variant enzyme engineering, the effect of multiple promoter driven Cas9, and gRNA efficient optimization and expression strategies. It also briefly introduces the optimization strategies of the CRISPR/Cas12a system and the application of BE and PE precision editing. These strategies are beneficial for the further development and optimization of gene editing systems in the field of plant molecular breeding.
Keywords: CRISPR/Cas9; gene editing; plant molecular breeding; SgRNA; PAM CRISPR/Cas9; gene editing; plant molecular breeding; SgRNA; PAM

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MDPI and ACS Style

Zhou, J.; Luan, X.; Liu, Y.; Wang, L.; Wang, J.; Yang, S.; Liu, S.; Zhang, J.; Liu, H.; Yao, D. Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding. Plants 2023, 12, 1478. https://doi.org/10.3390/plants12071478

AMA Style

Zhou J, Luan X, Liu Y, Wang L, Wang J, Yang S, Liu S, Zhang J, Liu H, Yao D. Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding. Plants. 2023; 12(7):1478. https://doi.org/10.3390/plants12071478

Chicago/Turabian Style

Zhou, Junming, Xinchao Luan, Yixuan Liu, Lixue Wang, Jiaxin Wang, Songnan Yang, Shuying Liu, Jun Zhang, Huijing Liu, and Dan Yao. 2023. "Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding" Plants 12, no. 7: 1478. https://doi.org/10.3390/plants12071478

APA Style

Zhou, J., Luan, X., Liu, Y., Wang, L., Wang, J., Yang, S., Liu, S., Zhang, J., Liu, H., & Yao, D. (2023). Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding. Plants, 12(7), 1478. https://doi.org/10.3390/plants12071478

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