Genetic expression box and construction method and application thereof

Abstract

The invention relates to a genetic expression box, and a construction method and application of the genetic expression box. The genetic expression box comprises a neuron specific promoter, a target gene, a target sequence of gamma-aminobutyric acid cholinergic neuron specificity miRNA, and a terminator which are connected in sequence. The target sequence of the gamma-aminobutyric acid cholinergic neuron specificity miRNA is added to the tail end of the target gene of the genetic expression box, due to the fact that a gamma-aminobutyric acid cholinergic neuron contains specific miRNA, the miRNA leads the genetic expression box to keep silent in the gamma-aminobutyric acid cholinergic neuron, and the target gene can not be expressed. A serotonin cholinergic neuron contains no miRNA, is not affected by gene silencing, and can normally express the target gene, thereby being stronger in specificity. Meanwhile, the specific promoter of the entire neuron is adopted, and the expression efficiency is high when the target gene is promoted in the neuron. A recombinant expression vector constructed by utilizing the genetic expression box or the genetic expression box can be widely applied in the research field of neural circuits related to the serotonin cholinergic neuron.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a gene expression cassette and its construction method and application. Background technique [0002] The use of cell-specific promoters or enhancers to achieve selective expression of foreign genes has been widely used in bioengineering fields such as gene therapy and optogenetics. But traditional technology also has many shortcomings, as follows: [0003] 1). Carrier capacity limitation [0004] The transgenic vectors used in neuroscience research are mainly recombinant adeno-associated virus and recombinant lentivirus. Adeno-associated virus is allowed to carry a maximum foreign gene of about 5000 base pairs, and lentiviral vector is allowed to carry a maximum foreign gene of about 7000 base pairs. The application of this technology is limited because the sequence of some specific promoters is too large, and the length of the target gene exceeds the maximum capacity of the cur...

AI Technical Summary

Problems solved by technology

[0005] 2). Restriction of transcriptional activity

[0007] 3). Certain cell types do not have specific promoters

[0011] Therefore, traditional serotonergic neuron-specific gene expression cassettes generally have the problems of poor specificity and low expression efficiency

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