Photo courtesy of the research team
Photo courtesy of the research team Recently, Jiang Linjian from the College of Plant Protection of China Agricultural University and Li Huarong from Qingdao Qingyuan Compound Co.
Chinese Academy of Sciences The researchers said that this study has further deepened the knowledge and understanding of gene editing tools
The third largest application scenario
The third largest application scenario Gene editing technology represented by CRISPR/Cas9 can generate precise mutations in the genome, providing a revolutionary tool for animal and plant breeding
Corresponding author Jiang Linjian introduced that at present, the application of gene editing technology to accurately mutagen genes is mainly concentrated in two scenarios: one is gene knockout, which is currently the most common application of gene editing in breeding
In view of the fact that a large number of genes can be improved by increasing their expression, gene knock-out (that is, up-regulation of gene expression) is the third largest application scenario in parallel with gene knockout and base editing to improve gene function
The general technical route for up-regulating gene expression is to insert regulatory elements, such as strong promoters, enhancers, etc.
"Knock high" rice endogenous genes
"Knock high" rice endogenous genes Since the overexpression of transgenic rice PPO1 and HPPD can respectively bring resistance to the two patented herbicides (Chinese independent intellectual property rights), the core question of this research is how to integrate synthetic DNA (as a strong promoter) without integration.
Many studies have long discovered that when Cas9 cuts at different positions of a gene at the same time, it will cause genomic structural variation such as deletions, inversions, and duplications
Based on this, the research team found the highly expressed genes CP12 and Ubiquitin2 near PPO1 and HPPD, respectively, based on the rice transcriptome information, and used related designs to construct a dual-target CRISPR vector to transform rice calli on a large scale.
In the successfully edited rice plants, the CP12 gene promoter drives the expression of the PPO1 gene, and the Ubiquitin2 gene promoter drives the expression of the HPPD gene, which greatly "knocks up" the endogenous PPO1 and HPPD genes in rice, making the rice plants perform.
The author stated that herbicide-resistant varieties combined with highly effective patented herbicides are expected to provide a more efficient solution for weed control in rice fields
It is expected to become a new field of gene editing technology
It is expected to become a new field of gene editing technology For a long time, Cas9 has been used as a gene knockout tool, and point mutations and up-regulation of gene expression usually require the insertion of artificial DNA templates.
Previously, the team developed a "circular target" base editing strategy.
Jiang Linjian said that this means that no artificial DNA template is needed.
With the development of long-range sequencing technology, it has become possible to compare the differences between different varieties at the genome level, and "pan-genomics" is in the ascendant
Therefore, the author believes that the structural variation of the genome is an important driving force for species evolution and a common phenomenon in nature
.
To create the expected structural variation through gene editing technology is based on nature as a teacher
.
Changing the expression pattern of genes is an important aspect of genetic improvement in animals and plants, but for a long time it has mainly been achieved by inserting artificial DNA
.
This research provides a universal method for changing gene expression patterns (especially knocking high genes) without inserting artificial DNA.
It is expected to open up a new field of application of gene editing technology in animal and plant breeding
.
(Source: China Science News Wang Fang and He Zhiyong)
Related paper information: https://doi.
org/10.
1038/s41477-021-01019-4
https://doi.
org/10.
1038/s41477-021-01019-4
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