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Construction method and application of sgRNA for knocking out human aldh2 gene, aldh2 gene deletion cell line

A gene deletion and construction method technology, applied in the field of genetic engineering, can solve problems such as expression recovery, and achieve the effect of improving incomplete silencing or inability to silence gene expression

Active Publication Date: 2020-10-16
SUN YAT SEN UNIV
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Problems solved by technology

[0006] Among the existing experimental technologies, the most similar to the CRISPR / Cas9 system is the siRNA-targeted gene silencing technology siRNA refers to the phenomenon of gene silencing induced by highly conserved double-stranded RNA in the evolution process of organisms, which affects a series of biological processes and function, but under the induction of drugs, etc., the expression of the silenced gene will appear to recover

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  • Construction method and application of sgRNA for knocking out human aldh2 gene, aldh2 gene deletion cell line
  • Construction method and application of sgRNA for knocking out human aldh2 gene, aldh2 gene deletion cell line
  • Construction method and application of sgRNA for knocking out human aldh2 gene, aldh2 gene deletion cell line

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Embodiment Construction

[0032] The following description is a preferred embodiment of the present invention, it should be pointed out that for those skilled in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications are also considered Be the protection scope of the present invention.

[0033] Unless otherwise specified in the examples of the present invention, all reagents and consumables used are commercially available.

[0034] The technical solution of the present invention is combined figure 1 The shown experimental design flow chart is realized through the following embodiments:

[0035] (1) sgRNA design:

[0036] The sgRNA sequences were designed for the second, fourth and fifth exons (Exon2, Exon4, Exon5) of the ALDH2 gene, respectively.

[0037] The specific grouping and naming of the three groups of designed and synthesized sgRNA sequences targeting the second, fourth and fifth ...

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Abstract

The invention provides an sg RNA sequence used for knocking out a human ALDH2 gene;,a target DNA sequence of the sg RNA is at least one of sequences as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3. The invention further provides a method for knocking out a human hepatoma carcinoma cell ALDH2 gene; the method utilizes a CRISPR / Cas system to modify the ALDH2 gene in a human hepatoma carcinoma cell. The invention further provides two ALDH2 gene deletion cell strains; ALDH2 participates in an important metabolic function of a body. The ALDH2 gene deletion cell strains provided by the invention provide an effective platform for metabolism study, in vivo, of exogenous chemicals or exogenous poisons, so that powerful means are provided for researching chronic diseases (such as alcoholic liver diseases and diabetes) as well as tumor-associated diseases.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a gRNA sequence for knocking out the human ALDH2 gene, a method for constructing an ALDH2 gene-deficient cell line and an application thereof. Background technique [0002] ALDH2 is a very important subtype of acetaldehyde dehydrogenase, which is located on chromosome 12 (12q24.2) in the human body. Its main function is to oxidize acetaldehyde into acetic acid, and it participates in the metabolism of acetaldehyde, a secondary metabolite of alcohol, in the body. Metabolic and detoxification processes, promoting alcohol metabolism in the liver, are thought to be closely related to alcohol metabolism and alcohol dependence in Asians. [0003] The lentiviral system is one of the methods currently used to mediate target sequences into target cells. It is a method developed on the basis of HIV. It overcomes the defect that some viral systems can only infect dividing cells. The lenti...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/90C12N5/10C12N9/22C12N15/867
CPCC12N9/22C12N15/113C12N15/86C12N15/907C12N2310/10C12N2320/11C12N2740/15043
Inventor 王庆王飞郭涛李若碧江红梅王婷曾妮
Owner SUN YAT SEN UNIV
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