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A kind of sgRNA guide sequence and its application of specific targeting mouse gdf5 gene

A species-specific, mouse-based technology, applied in the fields of medical genetics and molecular biology, can solve problems such as the lack of sgRNA guide sequences, and achieve the effects of facilitating drug screening of positively transfected cells, efficient gene editing efficiency, and high sensitivity

Active Publication Date: 2021-04-06
GUILIN MEDICAL UNIVERSITY
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Problems solved by technology

However, there is currently no sgRNA guide sequence for the mouse Gdf5 gene and no method to edit the gene

Method used

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  • A kind of sgRNA guide sequence and its application of specific targeting mouse gdf5 gene
  • A kind of sgRNA guide sequence and its application of specific targeting mouse gdf5 gene
  • A kind of sgRNA guide sequence and its application of specific targeting mouse gdf5 gene

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Embodiment Construction

[0076] 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.

[0077] 1. Screening of the pathogenic mutation site of the human Gdf5 gene, and confirming that glycine (G for short) at position 70 of the human Gdf5 gene is a pseudo-mutation site.

[0078] SYM1 is caused by mutations in the GDF5 gene, and two pathogenic mutations were detected by screening 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 mutation situation of the human GDF5 gene, and screens 164 mutation sites altogether (as shown in Table 2 and figure 1 shown). Then, the ClinVar database was used to search the pathogenicity of GDF5 missense mutations, and a total of 3 mutation sites were retrieved (as shown in Tabl...

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Abstract

The invention discloses a sgRNA guide sequence specifically targeting mouse Gdf5 gene and application thereof, belonging to the technical fields of medical genetics and molecular biology. The nucleotide sequence corresponding to the sgRNA is the sequence shown in SEQ ID NO.1. The present invention also discloses a method for editing the mouse Gdf5 gene by using the above-mentioned sgRNA guide sequence specifically targeting the mouse Gdf5 gene. The sgRNA guide sequence of the present invention can mediate the Cas9 protein to efficiently cut target DNA, and then be used to edit the mouse Gdf5 gene and affect the function of the protein encoded by the mouse Gdf5 gene. The sgRNA guide sequence of the present invention can realize high-efficiency targeting through the CRISPR / Cas9 system, and the efficiency is 100%.

Description

technical field [0001] The invention relates to a sgRNA guide sequence specifically targeting mouse Gdf5 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 / Cas...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/85C12N15/10C12N15/12C12Q1/02
CPCC07K14/51C12N15/102C12N15/1136C12N15/85C12N2503/02C12N2800/107C12N2310/20G01N33/5008G01N2500/10
Inventor 于鸿浩肖福英韦日明岳鹏鹏王添贤
Owner GUILIN MEDICAL UNIVERSITY
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