A kind of capsicum crispr-cas9 gene editing method and its application

Abstract

The invention discloses a capsicum CRISPR / Cas9 gene editing method and application thereof, belonging to the technical field of gene editing. Firstly, the sgRNA primers MMD1CaCRISPR-Cas9-F and MMD1CaCRISPR-Cas9-R were designed to construct the pepper gene editing vector; the plant pollen nano-magnetic transformation technology was used to mediate the transformation, and the gene editing of pepper was realized. The present invention uses plant pollen nano-magnetic transformation technology as the transformation method of gene editing in pepper for the first time, and obtains single-gene knockout mutant plants through CRISPR / Cas9 gene editing technology, and the mutation rate reaches 60%; for subsequent multi-gene knockout, gene Fragment insertion, replacement, etc. lay the technical foundation.

Description

A kind of pepper CRISPR-Cas9 gene editing method and its application technical field The invention belongs to the technical field of plant genetic engineering, be specifically related to a kind of pepper CRISPR-Cas9 gene editing method law and its applications. Background technique Transgenic technology plays a very important role in cultivating new plant varieties, and in traditional plant new products, cultivating all It needs to go through a tedious and long-term tissue culture process, which seriously affects the genetic transformation process. Genome-specific editing (site-specific genome editing) technology refers to the genome level Genetic manipulation techniques for site-directed modification of DNA sequences. It is widely used in genome modification, gene function research and new transgenic products It has great application value in cultivation and other aspects. However, the development of genome-directed editing technology has been slow, until By t...

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Problems solved by technology

In terms of site-specific editing of plant genomes, the CRISPR / Cas9 system has also been rapidly applied to dicotyledonous plants such as Arabidopsis and tobacco and monocotyledonous plants such as rice, sorghum, wheat, and corn, and has achieved a high rate of induced mutations and possible Stably inherited genome-edited plants; however, no gene-editing method for peppers has been reported

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