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Single guide RNA (sgRNA) guiding sequence specifically targeting mouse Galt gene and application of sgRNA guiding sequence

A species-specific, mouse-based technology, applied in the fields of medical genetics and molecular biology, can solve the problem of no sgRNA guide sequence knockout, etc., and achieve the effect of efficient screening, high concentration, and increased quantity

Active Publication Date: 2020-02-11
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 Galt gene and a method for knocking out the gene

Method used

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  • Single guide RNA (sgRNA) guiding sequence specifically targeting mouse Galt gene and application of sgRNA guiding sequence
  • Single guide RNA (sgRNA) guiding sequence specifically targeting mouse Galt gene and application of sgRNA guiding sequence
  • Single guide RNA (sgRNA) guiding sequence specifically targeting mouse Galt gene and application of sgRNA guiding sequence

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Embodiment Construction

[0084] 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.

[0085] 1. Screening of human GALT gene pathogenic mutation sites, and confirming that human GALT gene c.904+1G is a pseudo-mutation site.

[0086] Type I galactosemia is caused by mutations in the GALT gene. First, the inactivation mutation of the human GALT gene was screened using the ExAC database, and a total of 19 mutation sites were screened (as shown in Table 1 and figure 1 shown); in order to more extensively screen the pathogenic mutations of GALT, the present invention also utilizes the OMIM online database to retrieve 17 kinds of GALT mutations (as shown in Table 2); then, the present invention utilizes the ClinVar database to retrieve splicing site mutations The disease-causing conditions (as shown in Table 3). In view of th...

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Abstract

The invention discloses a single guide RNA (sgRNA) guiding sequence specifically targeting a mouse Galt gene and application of the sgRNA guiding sequence, and belongs to the technical field of medical genetics and molecular biology. The nucleotide sequence corresponding to sgRNA includes any one of the sequences shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. The invention further discloses amethod for editing the mouse Galt gene through the sgRNA guiding sequence specifically targeting the mouse Galt gene. The sgRNA guiding sequence can mediate a Cas9 protein to cut target DNA efficiently, and thus the sgRNA guiding sequence is used for editing the mouse Galt gene to affect the protein coding function of the mouse Galt gene. The sgRNA guiding sequence can mediate a CRISPR / Cas9 systemto achieve efficient target shooting with the efficiency of 100%.

Description

technical field [0001] The invention relates to a sgRNA guide sequence specifically targeting mouse Galt 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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IPC IPC(8): C12N15/113C12N15/85C12N15/90C12N5/10
CPCC07K14/47C12N15/113C12N15/907C12N2310/20
Inventor 于鸿浩岳鹏鹏付灿郭俊璠李勇
Owner GUILIN MEDICAL UNIVERSITY
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