A method for site-directed mutation of alfalfa gene using CRISPR/Cas9 system

A gene site-directed mutation, alfalfa technology, applied in the field of gene editing, can solve the problems of incomplete RNAi silencing, limited application, unstable characteristics, etc., and achieve the effect of thorough technology, high mutation efficiency, and accelerated breeding.
CN108866093BActive Publication Date: 2021-07-09GUANGDONG SANJIE HERBAGE BIOTECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
GUANGDONG SANJIE HERBAGE BIOTECH CO LTD
Publication Date
2021-07-09

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Abstract

The invention discloses a method for site-directed mutation of alfalfa gene by using CRISPR / Cas9 system. The present invention uses CRISPR / Cas9 technology in the cross-pollination autotetraploid plant alfalfa for the first time, and obtains mutant plants; for the first time, the MtU6 promoter is used to initiate sgRNA transcription in alfalfa to improve transcription efficiency; and natural mutation and traditional Compared with artificially induced mutations, the present invention accurately introduces mutations with higher mutation efficiency, and can achieve a mutation rate of 52%, of which the homozygous mutation rate reaches 12%. The invention directly mutates the gene coding sequence, which opens up a new field for gene editing in alfalfa, and lays the foundation for subsequent simultaneous silencing of multiple genes, deletion of large chromosome fragments, precise insertion of foreign genes into target sites, gene regulation, etc. technical basis.
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Description

technical field

[0001] The invention belongs to the technical field of gene editing, and in particular relates to a method for site-directed mutation of alfalfa gene by using a CRISPR / Cas9 system. Background technique

[0002] Alfalfa (Medicago sativa.) is a widely planted legume forage, rich in high-quality dietary fiber, edible protein, multivitamins (including vitamin B, vitamin C, vitamin E, etc.), a variety of beneficial minerals, saponins, flavonoids Bioactive components such as carotenoids, carotenoids, phenolic acids, etc., have high protein content and good palatability, which are of great significance for improving the feed level of livestock. In addition to being used as a forage crop, the well-developed root system of alfalfa plays an important role in the prevention and control of soil erosion and environmental governance. In addition, alfalfa also plays an important role in improving soil fertility because of the symbiosis of a large number of nitrogen-fixing ...

Claims

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