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Method for relieving miRNA inhibition function to promote target gene expression

A target gene and function technology, applied in genetic engineering, recombinant DNA technology, biochemical equipment and methods, etc., can solve problems such as few researches, and achieve the effect of simple operation, high efficiency, and low operating cost

Active Publication Date: 2019-08-23
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

However, there are few studies on the use of CRISPR-Cas9 to edit the miRNA complementary site of the target gene to release the repressive function of the miRNA.

Method used

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  • Method for relieving miRNA inhibition function to promote target gene expression
  • Method for relieving miRNA inhibition function to promote target gene expression
  • Method for relieving miRNA inhibition function to promote target gene expression

Examples

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Embodiment 1

[0035] Example 1: Using the CRISPR / Cas9 system to edit the target sequence of OsGRF4 that is inhibited by miRNA396 recognition to improve the grain shape of rice

[0036] Rice (Oryza sativa L.) is one of the most important staple food crops, feeding almost half of the world's population. The size of rice grain shape determines the yield per unit area of ​​rice and is also one of the main factors affecting grain production. OsGRF4 is a key gene regulating rice grain shape, and the expression of this gene can promote the increase of rice grain shape and increase rice yield. Previous studies have shown that miR396 can bind to and inhibit the transcriptional regulation of OsGRF 4, thereby making rice grains smaller and affecting rice yield. Based on the above background, using the strategies provided by the present invention, we use gene editing to destroy the target sequence of miR396 and screen for mutants that do not affect the function of OsGRF4 protein. By thus disrupting t...

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Abstract

The invention discloses a strategy for relieving a miRNA inhibition function to promote target gene expression, and provides a specific implementation method based on a genome editing technology. A gene editing technology is used for editing the recognition sequence of a miRNA target gene, base substitution mutation and amino acid deletion mutation with normal target gene functions are screened, the recognition sequence destroys mutants not inhibited by upstream miRNA, and the simple and efficient method is provided for promoting the functional research and functional application of the targetgene.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for releasing miRNA inhibition function and promoting target gene expression. Background technique [0002] MiRNA is a class of single-stranded RNA, usually 20-24nt, which plays an important post-transcriptional regulatory role in plants and animals. Small RNAs have been shown to be involved in many biological processes, including plant growth, development, stress responses, and hormone signaling. Binds to targeted mRNAs through a highly specific complementary base-pairing mechanism and mediates post-transcriptional gene silencing. Previous studies have shown that if there is an SNP in the target sequence recognized by miRNA on the target gene, it will destroy the recognition of miRNA and the inhibitory function of the target gene, thereby affecting the corresponding phenotype. It can be seen that interfering with the regulatory function of miRNA and its target gene may be...

Claims

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Application Information

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IPC IPC(8): C12N15/82
CPCC12N15/8216
Inventor 王锋林雅容朱义旺范美英
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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