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SgRNA guide sequence of specific targeting mouse Idua gene and application thereof

A kind of specific, mouse technology, applied in the fields of medical genetics and molecular biology, can solve the problem of no sgRNA guide sequence editing and other problems, achieve the effect of efficient screening, high concentration, and improve cell transfection efficiency

Active Publication Date: 2020-03-17
GUILIN MEDICAL UNIVERSITY
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  • Abstract
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Problems solved by technology

However, there is currently no sgRNA guide sequence for the mouse Idua gene and no method for editing the gene

Method used

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  • SgRNA guide sequence of specific targeting mouse Idua gene and application thereof
  • SgRNA guide sequence of specific targeting mouse Idua gene and application thereof
  • SgRNA guide sequence of specific targeting mouse Idua gene and application thereof

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Embodiment Construction

[0080] The principles and features of the present invention are described below in conjunction with specific drawings, and examples are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0081] 1. Screening of human IDUA gene pathogenic mutation sites, and confirming that glutamine (Q for short) at position 70 of human IDUA gene is a pseudo-mutation site

[0082] Mucopolysaccharidosis type I is caused by mutations in the IDUA gene, and 19 pathogenic mutations were screened out using the OMIM online database (as shown in Table 1). Due to the limited mutation sites included in OMIM, the present invention also utilizes the ExAC database to screen the function inactivation mutation of the human IDUA gene, and screens 19 mutation sites altogether (as shown in Table 2 and figure 1 shown). Then use the ClinVar database to retrieve the nonsense mutations of IDUA pathogenic sites, and a total of 14 mutation sites were retrie...

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Abstract

The invention, which belongs to the technical field of medical genetics and molecular biology, discloses an sgRNA guide sequence of a specific targeting mouse Idua gene and application thereof. The nucleotide sequence corresponding to the sgRNA is any one of SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3. In addition, the invention also discloses a method for editing the mouse Idua gene by using thesgRNA guide sequence of the specific target mouse Idua gene. According to the sgRNA guide sequence disclosed by the invention, Cas9 protein can be mediated and the target DNA can be cut efficiently, so that the mouse Idua gene is edited to affect the protein encoding function of the mouse Idua gene. The efficient targeting can be realized by using the CRISPR / Cas9 system and the efficiencies are respectively 100%, 83.33% and 100%.

Description

technical field [0001] The invention relates to a sgRNA guide sequence specifically targeting the mouse Idua gene and its application, 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 a 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 / C...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/85C12N15/90C12N5/10
CPCC07K14/47C12N15/113C12N15/907C12N2310/20
Inventor 于鸿浩岳鹏鹏付灿韦日明郭俊璠
Owner GUILIN MEDICAL UNIVERSITY
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