Multi-gene expression and silencing system controlled by heat shock protein gene promoter and tetracycline gene promoter

Abstract

The invention discloses a multi-gene expression and silencing system controlled by a heat shock protein gene promoter and a tetracycline gene promoter. The heat shock protein gene promoter HSPP and the tetracycline gene promoter TETp in the same plasmid system do not influence each other and independently control expression of downstream target genes or shRNAs (small hairpin ribonucleic acids) respectively, expression of single gene or shRNA can be controlled manually, and co-expression of two genes or shRNAs can be also controlled manually. The multi-gene expression and silencing system provides conditions for multi-gene specificity coordination control, solves the problem of low expression efficiency caused by gene co-expression implemented with a multi-plasmid cotransfection method at present, adopts a lentivirus carrying system to realize efficient expression of selective genes or shRNAs in cells having difficulty in transfection, meets various experimental requirements, provides a multi-gene target control platform, and provides more possibilities for gene therapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multi-gene expression and silencing system controlled by a heat shock protein gene promoter and a tetracycline gene promoter. Background technique [0002] When studying some specific genes such as genes closely related to cell growth, occurrence, and development, the inability to finely regulate the expression level of the target gene may affect the growth and proliferation of normal cells at the same time, so the regulatable and inducible system is considered to be the future An important development direction of gene therapy. Inducible expression vectors can regulate gene expression through inducible promoters, and then realize the timed and quantitative induced expression of target genes. The currently applied induction systems include cre2loxP system, tetracycline induction system, ecdysone induction system, heat shock induction system and lactose induction system. Wherein, ...

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Problems solved by technology

However, the current regulatable system has the following main defects: (1) Leaky expression, that is, there are different degrees of expression of the target gene under the state of regulation and inhibition, forming background noise. Since the mammalian gene system is complex, multi-level, Multi-pathway loops, basal expression is inevitable; (2) Among the genes for tumor treatment, single treatment often does not achieve a good therapeutic effect, while multi-target gene therapy can achieve better curative effect, the traditional control system Both are single induction control systems, which cannot realize the controllable control of multi-gene targets. Traditional multi-gene expression often adopts the method of plasmid co-transfection, and co-transfection leads to low gene expression efficiency, which cannot well meet the experimental requirements; (3) The traditional single induction control system cannot meet the needs of experiments when two genes need to be coordinated and controlled at the same time

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