Method for reducing rice amylose content through gene editing and special sgRNA

A technology of amylose content and gene editing, applied in the field of method and its special sgRNA, can solve instability and other problems, and achieve the effect of reducing amylose content

Active Publication Date: 2020-01-21
YUAN LONGPING HIGH TECH AGRI CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Amylose is soluble in water but unstable

Method used

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  • Method for reducing rice amylose content through gene editing and special sgRNA
  • Method for reducing rice amylose content through gene editing and special sgRNA
  • Method for reducing rice amylose content through gene editing and special sgRNA

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Embodiment Construction

[0022] The technical solution of the present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0023] The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0024] 1. Construction of gene editing vector

[0025] In order to obtain plants with reduced amylose content in 638S and Huazhan, the target site was designed according to the recognition characteristics of the CRISPR / Cas9 system, and a 20bp sgRNA target sequence ATCCACAACATTCCTACCA was designed in the exon 7 region of the Waxy gene, and the PAM sequence GGG was located in the target At the 3' end of the sequence, the CRISPR / Cas9 recombinant vector was constructed by double digestion with KpnI and HindIII ( figure 1 A), the recombinant vector was transformed into Agrobacterium EHA105 by electroporation. Using the GUS reporter gene on the recombinant vector, design specific detection...

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Abstract

The present invention discloses a method for reducing rice amylose content through gene editing and a special sgRNA, and belongs to the field of biotechnology. The method comprises the following steps: expression of a Waxy gene in rice is inhibited; and the Waxy gene is the gene encoding Waxy protein. A CRISPR/Cas9 technology is utilized to edit the rice Waxy gene on site. By causing frameshift mutations and knocking out the rice Waxy gene, a new generation of rice germplasm with significantly reduced amylose content is obtained. Compared with a wild type, the obtained Waxy site-edited strainis significantly lowered in amylose content.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for reducing rice amylose content through gene editing and a special sgRNA thereof. Background technique [0002] The main component of rice endosperm is starch, which generally accounts for 76% of the dry weight of rice flour, and exists in the endosperm in the form of irregular polygonal granules. Starch is divided into two types: amylose and amylopectin, both of which are polysaccharides combined with glucose and coated with a small amount of fat and protein. The amylose content of waxy rice is less than 2%, and the amylose content of non-waxy japonica rice varieties is generally lower than that of indica rice varieties. [0003] Amylose in rice endosperm is mainly catalyzed and synthesized by granule bound starch synthase I (GBSS I) encoded by rice waxy gene (Waxy), which is the main gene controlling amylose synthesis. The structure and function of Wx gene have been exte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82C12N15/29C07K14/415A01H5/00A01H6/46
CPCC12N15/113C12N15/8218C12N15/8245C07K14/415C12N2310/20
Inventor 杨远柱周延彪赵新辉唐倩莹唐晓丹廖雪
Owner YUAN LONGPING HIGH TECH AGRI CO LTD
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