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A set of vector system for rice epigenome methylation/demethylation

A demethylation and genome technology, applied in the direction of introducing foreign genetic material by using vectors, genetic engineering, plant gene improvement, etc., can solve problems such as developmental defects, unclear methylation, and gene expression changes

Pending Publication Date: 2020-08-21
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the generation of specific DNA methylation patterns is important for these processes, how methylation is regulated at each locus is unclear
Previous studies have found that DNA methylation of plant gene promoters sometimes leads to transcriptional repression, and loss of DNA methylation in methyltransferase mutants leads to altered gene expression and severe developmental defects

Method used

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  • A set of vector system for rice epigenome methylation/demethylation
  • A set of vector system for rice epigenome methylation/demethylation
  • A set of vector system for rice epigenome methylation/demethylation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, the establishment of the carrier for rice epigenome methylation / demethylation

[0036] 1. Carrier Construction

[0037] The circular plasmid shown in SEQ ID No.1 was artificially synthesized.

[0038] The plasmid includes expression cassette 1 and expression cassette 2.

[0039] The expression cassette 1 is shown in the 2889-10115 positions from the 5' end of SEQ ID No. 1 in the sequence listing. Wherein, the nucleotide sequence of the rice Act1 promoter is shown in the 2889-3731 position from the 5' end of SEQ ID No.1, and the nucleotide sequence of the TMVΩ enhancer is shown in the 3784-3784th position from the 5' end of SEQ ID No.1. 3839, the nucleotide sequence of dCas9 is shown in SEQ ID No.1 from 5' end 3856-7956, the nucleotide sequence of 1×HA tag is shown in SEQ ID No.1 from 5' end 7960 -shown at position 7986, the nucleotide sequence of 3×SV40 NLS is shown at positions 8005-8082 from the 5’ end of SEQ ID No.1, and the nucleotide sequence of 10...

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Abstract

The invention discloses a set of vector system for rice epigenome methylation / demethylation. The invention provides a set of vector system for rice epigenome demethylation. According to the system, DNA methylation variation is rapidly generated by randomly demethylating a rice genome. The system relates to the expression of rice OsALKBH DNA 6mA demethylase, resulting in a wide range of low methylation. The mutagenesis technique is of great significance to plants with important agricultural significance, which may result in differential expression of alleles that are generally silenced by DNA methylation, thereby revealing the application of previously hidden phenotypic variations.

Description

technical field [0001] The invention relates to the technical field of gene directional expression circuit synthesis, in particular to a set of vector systems for rice epigenome methylation / demethylation. Background technique [0002] Genetic engineering and synthetic biology have become important means of researching and improving crop traits. With the development and utilization of gene editing tools zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEs) and CRISPR / Cas9 system, the conditions for targeting and modifying specific genome sequences to study related gene functions have changed. Be very mature. A growing body of research is using gene editing tools for epigenetic modification as well as gene regulation. [0003] Rice is one of the main food crops in the world, and it is also a model plant for functional genomics research. Studies based on the rice genome have been relatively mature, however, research on epigenetic modifications th...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/53A01H5/00A01H6/46
CPCC12N9/0071C12N15/8216C12N15/8218C12Y114/11
Inventor 谷晓峰胡桂花
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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