sgRNA guide sequence of specific targeting mouse Tyr gene and application 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 sequences, achieve efficient screening, improve cell transfection efficiency, and achieve high-concentration effects

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

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Problems solved by technology

However, there is currently no sgRNA guide sequence for the mouse Tyr gene and no method to edit the gene

Method used

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

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Embodiment Construction

[0080] 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.

[0081] 1. Screening of the human Tyr gene pathogenic mutation site, confirming that the proline (referred to as Pro) site at position 81 of the human TYR gene is a pseudo-mutation site.

[0082] I albinism is caused by mutations in the TYR gene, and 38 pathogenic mutations were screened using the OMIM online database (as shown in Table 1, a total of 20 are listed). Due to the limited mutation sites included in OMIM, the present invention also utilizes the ExAC database to screen the mutation situation of the human TYR gene, and screens 283 mutation sites altogether (as shown in Table 2 and figure 1 Shown, Table 2 lists 20 kinds of mutations). Then, the ClinVar database was used to search for mutations at TYR pathogenic sites, and a tot...

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Abstract

The invention discloses an sgRNA guide sequence of a specific targeting mouse Tyr 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 Tyr gene by utilizing the sgRNA guidesequence of the specific targeting mouse Tyr 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 usedfor editing the Tyr gene of a mouse to influence the function of the Tyr gene encoding protein of the mouse. 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 Tyr 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 microorganisms by means of genetic engineering, thereby creating a gene editing targeting system that can be widely used in 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 system, unde...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85C12N15/90C12Q1/02
CPCC12N9/0059C12N15/1137C12N15/907C12N2503/02C12N2800/107C12Y110/03001C12N2310/20G01N33/5008G01N33/5044G01N2500/10
Inventor 岳鹏鹏于鸿浩付灿肖福英李勇
Owner GUILIN MEDICAL UNIVERSITY
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