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On December 8, 2022, the Institute of Crop Science, Chinese Academy of Agricultural Sciences/Sanya Nanfang Institute of Crop Precision Breeding Technology Innovation Team was invited to publish a review article online in the Journal of Integrative Plant Biology (JIPB), focusing on reviewing the development process and application of base editor and guide editor in crop improvement, and proposing the future research and development direction
of plant base editor and guide editor.
of plant base editor and guide editor.
The essence of crop variety improvement is the aggregation of excellent alleles, however, due to gene linkage effect, the introduction of excellent alleles in breeding practice often requires years of continuous hybridization and backcrossing, which is laborious and time-consuming
.
CRISPR/Cas gene editing technology can carry out targeted deletion, replacement and insertion of target genes, and because of its advantages of simple operation, high editing efficiency and multi-target editing, it has shown broad application prospects
in gene function research and improvement of important agronomic traits of crops.
Usually, the difference between crop varieties is only the difference of a few bases or small fragments between alleles, and the use of genome precision editing technology can achieve fixed-point base replacement, small fragment replacement or insertion, and rapid, directional and efficient creation of new crop materials, and then cultivate new crop varieties
with excellent agronomic traits.
.
CRISPR/Cas gene editing technology can carry out targeted deletion, replacement and insertion of target genes, and because of its advantages of simple operation, high editing efficiency and multi-target editing, it has shown broad application prospects
in gene function research and improvement of important agronomic traits of crops.
Usually, the difference between crop varieties is only the difference of a few bases or small fragments between alleles, and the use of genome precision editing technology can achieve fixed-point base replacement, small fragment replacement or insertion, and rapid, directional and efficient creation of new crop materials, and then cultivate new crop varieties
with excellent agronomic traits.
At present, there are three main CRISPR/Cas-mediated genome precision editing technologies, including homologous recombinant technology (HDR), single-base editing technology and guided editing technology
.
HDR in plants remains challenging
due to the low frequency of CRISPR/Cas-mediated HDR in plant cells, especially in plant cells that cannot efficiently deliver sufficient repair templates near the double-strand break gap.
In contrast to HDR, single-base editing and guided editing techniques do not require the involvement
of double-stranded break notches and exogenous donor repair templates.
Single-base editing technology can precisely mutate one base pair into another in the target region
.
Guided editing techniques enable all types of single-base free conversion and reversal as well as site-directed substitution and insertion
of small fragments within the target area.
The development and utilization of single-base editing and guided editing technology have greatly expanded the scope and ability of precise editing of plant genomes, and provided important technical support
for the precise and targeted improvement of important agronomic traits of crops.
.
HDR in plants remains challenging
due to the low frequency of CRISPR/Cas-mediated HDR in plant cells, especially in plant cells that cannot efficiently deliver sufficient repair templates near the double-strand break gap.
In contrast to HDR, single-base editing and guided editing techniques do not require the involvement
of double-stranded break notches and exogenous donor repair templates.
Single-base editing technology can precisely mutate one base pair into another in the target region
.
Guided editing techniques enable all types of single-base free conversion and reversal as well as site-directed substitution and insertion
of small fragments within the target area.
The development and utilization of single-base editing and guided editing technology have greatly expanded the scope and ability of precise editing of plant genomes, and provided important technical support
for the precise and targeted improvement of important agronomic traits of crops.
This paper first reviews the latest progress in the development of various base editors and guide editors and their applications
in crop genetic improvement.
On this basis, this paper recommends several more efficient base editors (Figure 1) in crop gene function research and genetic improvement, such as evoFERNY-CBE base editor (Figure A), TadA9-ABE or hyTadA8e-ABE base editor (Figure B), as well as efficient boot editors such as PEmax (Figure D) and pegRNA carrying evopreQ1 (Figure E).
On this basis, combined with the proxy base editing and guided editing system, the efficiency
of plant single-base editing and guided editing can be further improved.
Finally, this paper puts forward the future optimization and improvement direction of plant base editor and guide editor, such as narrowing or expanding the editing window of base editor and developing base editor suitable for different purposes.
Develop a more efficient guide editor by introducing elements such as single-stranded annealed proteins; Development of technology systems for guided editing-mediated large-fragment allele replacement or insertion.
in crop genetic improvement.
On this basis, this paper recommends several more efficient base editors (Figure 1) in crop gene function research and genetic improvement, such as evoFERNY-CBE base editor (Figure A), TadA9-ABE or hyTadA8e-ABE base editor (Figure B), as well as efficient boot editors such as PEmax (Figure D) and pegRNA carrying evopreQ1 (Figure E).
On this basis, combined with the proxy base editing and guided editing system, the efficiency
of plant single-base editing and guided editing can be further improved.
Finally, this paper puts forward the future optimization and improvement direction of plant base editor and guide editor, such as narrowing or expanding the editing window of base editor and developing base editor suitable for different purposes.
Develop a more efficient guide editor by introducing elements such as single-stranded annealed proteins; Development of technology systems for guided editing-mediated large-fragment allele replacement or insertion.
Li Jingying, a postdoctoral fellow at the Institute of Science and Technology/Hainan Yazhou Bay Seed Laboratory, and Dr.
Zhang Chen, an assistant researcher, are the co-first authors of the paper, and researcher Xia Lanqin is the corresponding author
.
The research was supported
by the National Key Research and Development Program, the Southern Breeding and Breeding Project of the Chinese Academy of Agricultural Sciences, the Hainan Yazhou Bay Seed Laboratory and the Key Laboratory of Gene Editing Technology (Hainan) of the Ministry of Agriculture and Rural Affairs of China.
Zhang Chen, an assistant researcher, are the co-first authors of the paper, and researcher Xia Lanqin is the corresponding author
.
The research was supported
by the National Key Research and Development Program, the Southern Breeding and Breeding Project of the Chinese Academy of Agricultural Sciences, the Hainan Yazhou Bay Seed Laboratory and the Key Laboratory of Gene Editing Technology (Hainan) of the Ministry of Agriculture and Rural Affairs of China.
Article link: https://doi.
org/10.
1111/jipb.
13425
org/10.
1111/jipb.
13425
