Separated encoded IFNLR1 mutant nucleic acid and application thereof

A mutant and nucleic acid technology, applied in the field of susceptibility to non-syndromic autosomal dominant genetics, can solve problems such as unknown disease-causing gene loci, and achieve the effect of broadening the scope of clinical diagnosis and detection/screening

Active Publication Date: 2019-06-28
GENERAL HOSPITAL OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, 36 genes have been reported to be related to ADNSHL, but there are still a considerable number of unknown pathogenic loci

Method used

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  • Separated encoded IFNLR1 mutant nucleic acid and application thereof
  • Separated encoded IFNLR1 mutant nucleic acid and application thereof
  • Separated encoded IFNLR1 mutant nucleic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1 Whole Exome Sequencing Determination of Disease-causing Genes and Mutation Sites

[0080] 1. Sample collection

[0081] The inventor collected a pedigree of Chinese Han hereditary deafness with morbidity for three consecutive generations (see the pedigree for the pedigree) figure 2 ), the family consists of 24 members, including 10 confirmed patients and 9 existing patients (diagnostic diagram see image 3 ), collected the peripheral blood samples of the family members, among which the collected blood samples were 9 cases (II:2, II:3, II:4, II:5, II:6, III:3, III:5 , III:6, III:7); collected 14 cases of phenotype normal human blood samples (I:2, II:1, II:7, II:8, II:9, II:10, II:11 , II:12, III:1, III:2, III:4, III:8, III:9, III:10). 2 mL of peripheral blood samples were collected from each sample, added with EDTA for anticoagulation, and stored at -80°C.

[0082] in, figure 2 The pedigree chart is shown, where ○ indicates a normal female, □ indicates a...

Embodiment 2

[0108] Example 2 Sanger method sequencing to verify the pathogenic mutation of ADNSHL disease

[0109] The IFNLR1 gene obtained in the detection of Example 1 was detected respectively, primers were designed for the sequence of the mutation site involved in the above gene, and then the relevant sequence of the above mutation was obtained by PCR amplification, product purification and sequencing, and the determined sequence was determined according to the result Whether it is mutant or wild type, whether it is heterozygous mutation or homozygous mutation, and whether the sequence and phenotype are co-segregated in the family to verify the correlation between the above genes and ADNSHL.

[0110] 1. DNA extraction

[0111] Collect 10ml of peripheral venous blood, use OMEGA Blood DNA Midi Kit whole blood DNA extraction kit to extract genomic DNA, and use spectrophotometer and gel electrophoresis to measure the concentration and purity of DNA and dilute to 200ng / μl;

[0112] 2. Pri...

Embodiment 3

[0128] IFNLR1 gene expresses Interferon lambda receptor 1 (Interferon lambda receptor 1) protein, which is related to the recognition of cytokines and interferon in the extracellular environment and intracellular signal transduction. , the spiral ganglion and the vestibular epithelium of the inner ear. Its functions include antiviral activity, antiproliferation, anticancer activity, molecular expression of MHC (major histocompatibility complex) complex I / II, and immune response. So far, the role of IFNLR1 in the auditory system has not been discovered.

[0129] In order to further study the function of IFNLR1 in the auditory system, the present invention uses a mouse model to study the expression and distribution of IFNLR1 in the inner ear of the mouse by means of fluorescent labeling, and finds that the IFNLR1 gene is widely expressed in the cochlea and the spiral organ.

[0130] Using the zebrafish model, the IFNLR1 gene mutant zebrafish was compared with the wild-type zebr...

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Abstract

The invention relates to a gene mutant and application thereof, in particular to application of a reagent for detecting a mutation site for preparing and screening out a biological sample easily infected with nonsyndromic autosomal dominant hereditary hearing loss, separated encoded IFNLR1 mutant nucleic acid, separated polypeptide, a system for screening out the biological sample easily infectedwith nonsyndromic autosomal dominant hereditary hearing loss, and a reagent kit, a construction body and reconstitution cells for screening out the biological sample with the nonsyndromic autosomal dominant hereditary hearing loss. Compared with a wild IFNLR1 gene, the separated encoded IFNLR1 mutant nucleic acid has c.296G>A mutation. By detecting whether or not the novel mutant exists in the biological sample, whether or not the biological sample is easily infected with the ADNSHL syndrome can be effectively detected.

Description

technical field [0001] The present invention relates to gene mutant and its application. Specifically, the present invention relates to isolated nucleic acids encoding IFNLR1 mutants and applications thereof, more specifically, to isolated nucleic acids encoding IFNLR1 mutants, isolated polypeptides, screening for non-syndromic autosomal dominant deafness A biological sample system, a kit for screening biological samples susceptible to non-syndromic autosomal dominant deafness, a construct, a recombinant cell, and a method for constructing a drug screening model. Background technique [0002] Non-syndromic deafness is a kind of hereditary deafness, which means that deafness is the only symptom of the individual with no other genetic damage and organ dysfunction, accounting for about 70% of hereditary deafness. Depending on the mode of inheritance, it is usually divided into autosomal dominant (DFNA), autosomal recessive (DFNB), sex-linked (sex-linked) and mitochondrial (mit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/47
Inventor 高雪戴朴袁永一林琼芬管李萍张建国党孝
Owner GENERAL HOSPITAL OF PLA
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