Multigene 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 heat shock protein gene promoter and tetracycline gene promoter. The expression of the downstream target gene or shRNA can be artificially controlled either the expression of a single gene or shRNA, or the co-expression of two genes or shRNA. The present invention provides conditions for multi-gene specific coordinated control, solves the problem of low expression efficiency brought about by the multi-plasmid co-transfection method to achieve gene co-expression, and adopts the lentivirus carrying system to be able to transfect cells that are difficult to transfect The high-efficiency expression of selective genes or shRNAs can be achieved in the medium, which meets the needs of various experiments, and at the same time provides a multi-gene target control platform, providing 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 (leaky expression), that is, there are different degrees of expression of the target gene under the state of regulation and inhibition, forming background noise. (2) Among the genes for tumor treatment, single treatment often fails to achieve a good therapeutic effect, while multi-target gene therapy can achieve better curative effect, and traditional control systems cannot It is a single induction control system, 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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