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Skeletal Muscle Cell Efficient Specific Promoter and Its Application

A high-efficiency start-up and promoter technology, which can be used in the fields of genetically active ingredients, animal/human peptides, and introduction of foreign genetic material using vectors, which can solve the problems of unsustainable expression of foreign genes and restricted applications.

Active Publication Date: 2019-08-06
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the efficiency of these natural muscle-specific promoters to express foreign genes is usually much lower than that of viral promoters, such as CMV promoters, thus limiting their application in muscle tissue
[0006] There have been some studies on synthetic promoters. The SPc5-12 promoter is a strong muscle-specific synthetic promoter, which is composed of SRE elements (serum response element), muscle cell enhancer factors 1 and 2 (MEF-1 and MEF-2) and transcription enhancer factor (TEF-1) 4 kinds of regulatory sequences randomly combined and screened, but this promoter cannot express foreign genes continuously

Method used

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  • Skeletal Muscle Cell Efficient Specific Promoter and Its Application
  • Skeletal Muscle Cell Efficient Specific Promoter and Its Application
  • Skeletal Muscle Cell Efficient Specific Promoter and Its Application

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: Preparation of high-efficiency specific promoters for skeletal muscle cells

[0026] 1. Preparation method of high-efficiency promoter

[0027] 1. Synthetic regulatory elements

[0028] Complementary oligonucleotides of 19 kinds of regulatory elements were sent to Sangon Bioengineering (Shanghai) Co., Ltd. for synthesis separately. The oligonucleotide sequences are shown in Table 1.

[0029] Table 1 19 regulatory elements

[0030]

[0031] 2. Phosphorylation and annealing of synthetic small fragments

[0032] The complementary oligonucleotides of the 19 kinds of regulatory elements synthesized were respectively diluted, phosphorylated and annealed according to the synthesis instructions to obtain DNA fragments of each regulatory element. The phosphorylation and annealing reactions were carried out in a 30 μl reaction system, and the steps were as follows. First, the complementary oligonucleotide sequence of each element was diluted to 100 μM, and then c...

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Abstract

The invention discloses an efficient and specific promoter of skeletal muscle cells and application of the efficient and specific promoter. According to the efficient and specific promoter disclosed by the invention, the nucleotide sequence is shown as any one of SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. The invention further discloses a recombinant vector and / or recombinant bacterium containing the promoter and application thereof. The expression efficiency of the efficient and specific promoter of the skeletal muscle cells in in-vitro and in-vivo experiments is higher than that of CMV (Cytomegalovirus) and high expression of target genes can be realized; the promoter SP-301 has remarkable advantages in the aspects of strength, specificity and lasting time of promoter gene expression in skeletal muscles, and has a very great potential in a subsequent gene therapy field.

Description

technical field [0001] The invention relates to a high-efficiency specific promoter for skeletal muscle cells and its application. Background technique [0002] In gene therapy, intramuscular injection of a therapeutic gene is a simple and effective treatment. Skeletal muscle has the characteristics of wide distribution, large number, and rich blood vessels. Its muscle cells, especially striated muscle cells, also have the characteristics of large cell size and multinucleation, so that it has the advantages of large volume, large injection capacity and easy operation. Functional secretion Protein is easier to enter the blood circulation system to play a therapeutic role. In addition, based on the long life cycle of skeletal muscle, the simplicity and ease of intramuscular injection, and the ability to take in a large amount of foreign genes, and the recombinant therapeutic protein can be continuously expressed in muscle tissue for several months to years, skeletal muscle ha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/63A61K48/00
CPCA61K9/0019A61K48/005C07K14/47C12N15/63
Inventor 江明锋王刚刘益丽
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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