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Method for preparing retinopathy non-human mammal model and application of mammal model

A technology for non-human mammals and retinopathy, applied in the fields of molecular biology and biomedicine, can solve the problems of no animal models, etc., and achieve the effect of simple steps, high efficiency, and no interference of foreign genes

Inactive Publication Date: 2020-11-24
上海朗昇生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no relevant animal model report of CNGA1 gene knockout

Method used

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  • Method for preparing retinopathy non-human mammal model and application of mammal model
  • Method for preparing retinopathy non-human mammal model and application of mammal model
  • Method for preparing retinopathy non-human mammal model and application of mammal model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment C

[0052] Construction and Identification of Example CNGA1-KO Mouse Model

[0053] 1. Materials

[0054] The mouse strain used in this example is: C57BL / 6J, and the surrogate mother mouse strain is C57BL / 6J, which was purchased from Shanghai Slack Experimental Animal Co., Ltd., and the mice were randomly divided into a control group and an experimental group.

[0055] Various restriction endonucleases, T4 DNA ligase, Taq enzyme and PCR related reagents were purchased from NEB Company. Conventional chemical reagents were mainly purchased from Sigma and Shanghai Chemical Reagent Company. The gel recovery kit, the plasmid extraction kit were purchased from Qiagen, and the in vitro transcription kit was purchased from Invitrogen.

[0056] Primers, etc. were synthesized from Suzhou Jinweizhi Biotechnology Co., Ltd.

[0057]

[0058]

[0059] 2. Method

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Abstract

The invention relates to the technical field of molecular biology and biomedicine, in particular to a method for preparing a retinopathy non-human mammal model and an application of the mammal model.The method comprises the following steps: knocking out a partial sequence of a second exon of an animal CNGA1 gene by using a Crispr-Cas system, wherein the Crispr-Cas system comprises sgRNA and Cas9enzyme, and the sgRNA comprises sgRNA1 and sgRNA2 of which the corresponding target site sequences are shown as SEQ ID NO: 1 and SEQ ID NO: 2.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and biomedicine, in particular to a method for preparing a non-human mammalian model of retinopathy and its application. Background technique [0002] CNGA1 gene, full name cyclic nucleotide gated channel subunit alpha 1, also known as CNCG, CNG1, RP49, etc., in human tissues, this gene is located on p12 on chromosome 4, encodes two different subtypes of transcripts, and is widely expressed in the liver , also expressed in kidney, fat, skin, bladder and other tissues. CNGA1 (NCBI: NM_001142564.1), encoding cyclic nucleotide-gated channel alpha1 transcript 1, has 10 exons, and this transcript variant encodes a longer transcript. CNGA1 (NCBI: NM_000087.4), encoding cyclic nucleotide-gated channel alpha1 transcript 2 with 11 exons, differs in 5'UTR and coding sequence compared to transcript 1 , the resulting transcript has a shorter N-terminus than transcript 1. This gene encodes a memb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89C12N15/113C12N15/12A01K67/027A61K49/00
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/03A61K49/0008C07K14/47C12N15/113C12N15/89C12N2310/20
Inventor 汪枫桦
Owner 上海朗昇生物科技有限公司
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