Construction method and application of sg RNA and ALDH2 gene deletion cell strains used for knocking out human ALDH2 gene

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

Active Publication Date: 2017-12-22
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 sg RNA and ALDH2 gene deletion cell strains used for knocking out human ALDH2 gene
  • Construction method and application of sg RNA and ALDH2 gene deletion cell strains used for knocking out human ALDH2 gene
  • Construction method and application of sg RNA and ALDH2 gene deletion cell strains used for knocking out human ALDH2 gene

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Embodiment Construction

[0032] The following are the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications are also considered This is the protection scope of the present invention.

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

[0034] The technical scheme of the present invention combines figure 1 The experimental design flow chart shown is realized by the following embodiments:

[0035] (1) sgRNA design:

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

[0037] The three groups of sgRNA sequences designed and synthesized for the second, fourth and fifth exons of ALDH2 gene are grouped and named in Table 1:

[0038] Tab...

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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 present invention relates to the field of genetic engineering, in particular to a method for knocking out the gRNA sequence of the human ALDH2 gene, an ALDH2 gene deletion cell line and its application. Background technique [0002] ALDH2 is a very important subtype of acetaldehyde dehydrogenase. It is located on chromosome 12 (12q24.2) of the human body. Its main function is to oxidize acetaldehyde into acetic acid. It participates in the production of alcohol's secondary metabolite acetaldehyde in the body. The process of metabolism and detoxification, which promotes the metabolism of alcohol in the liver, is believed 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 the sequence of interest 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 lentiviral...

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Application Information

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Patent Type & Authority Applications(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汪红梅王婷曾妮
Owner SUN YAT SEN UNIV
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