sgRNA guide sequence of specific targeting mouse Gaa gene and application of thereof

A species-specific, mouse-based technology, applied in the fields of medical genetics and molecular biology, can solve problems such as lack of sgRNA guide sequence editing, and achieve the effects of easy cultivation, increased quantity, and high DNA transfection efficiency

Active Publication Date: 2020-03-06
GUILIN MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is currently no sgRNA guide sequen...

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  • sgRNA guide sequence of specific targeting mouse Gaa gene and application of thereof
  • sgRNA guide sequence of specific targeting mouse Gaa gene and application of thereof
  • sgRNA guide sequence of specific targeting mouse Gaa gene and application of thereof

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Embodiment Construction

[0079] The principles and features of the present invention will be described below in conjunction with specific drawings, and the examples given are only used to explain the present invention, not to limit the scope of the present invention.

[0080] 1. The pathogenic mutation site of human GAA gene was screened, and the human GAA p.Tyr609 site was identified as the pseudo-mutation site.

[0081] Pompe disease is caused by mutations in the GAA gene. First, the pathogenic mutations in the human GAA gene were screened using the OMIM database, and a total of 16 mutation sites were screened (as shown in Table 1); for wider screening The pathogenic mutation of GAA, the present invention also utilizes the ExAC online database to retrieve 46 kinds of GAA mutations (such as figure 1 And shown in table 2, table 2 lists 20 kinds of mutations altogether); Then, the present invention utilizes ClinVar database to search GAA base deletion mutation situation again, retrieves 83 kinds of mut...

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Abstract

The invention discloses an sgRNA guide sequence of a specific targeting mouse Gaa gene and an application thereof, which belong to the technical field of medical genetics and molecular biology. A nucleotide sequence corresponding to the sgRNA is any one of SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3. The invention also discloses a method for editing the mouse Gaa gene by using the sgRNA guide sequence of the specific targeting mouse Gaa gene. The sgRNA guide sequence disclosed by the invention can mediate Cas9 protein to efficiently cut target DNA, so that the sgRNA guide sequence is used forediting the mouse Gaa gene to influence the function of encoding protein of the mouse Gaa gene. According to the sgRNA guide sequence, efficient targeting can be achieved through a CRISPR/Cas9 system,and the efficiency is 100%.

Description

technical field [0001] The invention relates to a sgRNA guide sequence specifically targeting mouse Gaa gene and application thereof, belonging to the technical fields of medical genetics and molecular biology. Background technique [0002] The CRISPR / Cas system was first discovered in bacteria, and it performs acquired immune functions in eubacteria and archaea, which can resist the invasion of foreign viruses and plasmids. People have modified the CRISPR / Cas system of the microorganism itself by means of genetic engineering, thereby creating a targeting system that can be widely used in gene editing of higher organisms, the CRISPR / Cas9 system. Since the system came out in 2012, it has injected strong impetus into life science and biomedical research, and has become a research hotspot in recent years. Since 2013, researchers have successfully edited the genome in mammalian cells by using the DNA binding activity and endonuclease activity of CRISPR / Cas9. In the CRISPR / Cas9...

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Application Information

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IPC IPC(8): C12N15/113C12N15/85C12N15/90C12Q1/02
CPCC12N9/2408C12N15/1137C12N15/907C12N2503/02C12N2800/107C12Y302/0102C12N2310/20G01N33/5008G01N33/5044G01N2500/10
Inventor 于鸿浩付灿岳鹏鹏李勇高进涛
Owner GUILIN MEDICAL UNIVERSITY
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