Gene editing vector and gene editing method and application thereof

Abstract

The invention relates to the technical field of gene engineering, in particular to a gene editing vector and a gene editing method and application thereof. The gene editing vector comprises a sequence of potyvirus from a 5'-UTR region to a capsid protein region, gRNA, an insertion sequence and a 3'-UTR region which are arranged in sequence, and the insertion sequence comprises a nucleic acid sequence for coding the virus capsid protein of the potyvirus. According to the gene editing vector, a gene editing system is constructed through the potyvirus, so that a variety of crops, such as soybeans and potatoes, which are less involved in the prior art can be dip-dyed, and the object and range of gene editing are expanded. Meanwhile, the technical problem that the gRNA is difficult to generate in a conventional mode due to the fact that the virus of the potyvirus cannot generate subgenome RNA is solved, and the method has important significance in the technical field of genetic engineering.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a gene editing vector and its gene editing method and application. Background technique [0002] In many bacteria and most archaea, there is a defense system that can be used to resist foreign DNA (such as phage), called CRISPR / Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR-associated Cas9 endonuclease) system (Bhaya et al ., 2011). The CRISPR / Cas9 system consists of two basic components, one is the endonuclease Cas9 protein, and the other is gRNA (guide RNA). Under certain conditions, the gRNA can form a complex with the Cas9 protein, and through complementary base pairing, the gRNA can guide the Cas9 protein to the target DNA, and then the Cas9 protein can bind and cut the target double-stranded DNA, thereby causing the target DNA double-strand break ( double strand break). Intracellular DNA repair mechanisms repair damaged double-stranded...

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Problems solved by technology

However, the host range of TRV is relatively limited and cannot infect important crops such as barley, wheat, and corn

[0013] In summary, most of the current gene editing systems that use plant viruses to deliver guide RNA rely on Cas9 transgenic plants (except SYNV), and the range of hosts is still not wide enough, and the hosts that can be applied are still limited

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