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Recombinant collagen-mRNA (messenger ribonucleic acid) composite material and preparation method thereof

A composite material, recombinant collagen technology, applied in medical science, prosthesis, etc., can solve the problems of carcinogenic effect immune response, easy decomposition or diffusion, mRNA instability and so on

Inactive Publication Date: 2014-12-24
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Defects of viral vectors include endogenous recombination, oncogenic effects, and unexpected immune responses
Also, transplanting dividing cells runs the risk of tumor formation
The third method is to combine GF with biomaterial carriers, but the denaturation and explosive release of growth factors are still difficult problems (Journal of the Royal Society Interface, 2011, 8:153-170; Gene Therapy, 2002, 9:1647 -1652)
However, mRNA is unstable in tissues, easy to decompose or diffuse, and requires suitable scaffolding materials to provide a place for protein synthesis

Method used

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  • Recombinant collagen-mRNA (messenger ribonucleic acid) composite material and preparation method thereof
  • Recombinant collagen-mRNA (messenger ribonucleic acid) composite material and preparation method thereof
  • Recombinant collagen-mRNA (messenger ribonucleic acid) composite material and preparation method thereof

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Embodiment Construction

[0031] The technical solutions of the present invention will be further illustrated and described below through specific embodiments in conjunction with the accompanying drawings.

[0032] In the following specific embodiments, the green fluorescent protein eGFP is selected to simulate the specific protein factor

[0033] (1) Cloning the DNA sequence encoding the amino acid sequence of the collagen into the pET-MBP-TevH plasmid to construct the protein vector recombinant plasmid:

[0034] As shown in SEQ ID 1, the collagen is based on human type I collagen and contains an amino-terminal His 6 10 glycine-proline-proline (GPP) were added to the label, TEV enzyme cleavage site, and both ends to increase the stability of collagen. Two glycine-proline-cysteine ​​(GPC) were inserted into the collagen sequence to provide mRNA cross-linking sites, and the distance between the two cysteines was about 30nm.

[0035] As shown in SEQ ID 2, the DNA sequence was cloned into pUC57 plasmid ...

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Abstract

The invention discloses a recombinant collagen-mRNA (messenger ribonucleic acid) composite material. The composite material comprises mRNA for coding specific protein factors and a protein carrier loading the mRNA, wherein the protein carrier sequentially comprises a maltose-binding protein and a collagen from an amino terminal to a carboxyl terminal; the mRNA is in covalent linkage with cysteine in the glycine-proline-cysteine sequence of the collagen of the protein carrier through an amino group modifying the 3' terminal. The composite material is capable of continuously and controllably releasing the specific protein factors to a wound part.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and in particular relates to a recombinant collagen-mRNA composite material and a preparation method thereof. Background technique [0002] Tissue engineering is to rebuild damaged tissues by planting cells on biomaterial scaffolds or compounding growth factors with scaffolds and transplanting them into the body. A prominent problem with the use of autologous cells is the difficulty of harvesting sufficient numbers, especially when the patient is very old or seriously ill. Allogeneic cells are often used for skin tissue engineering, and this engineered tissue can secrete growth factors that promote tissue repair. Xenogeneic cells are commonly used in epithelial tissue engineering requiring keratinocytes, although they can strongly promote epidermal tissue growth, but there is a risk of viral infection. In addition, both allogeneic and xenogeneic cells are immunogenic and require im...

Claims

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Application Information

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IPC IPC(8): A61L27/54A61L27/24A61L27/22
Inventor 孙莉萍约纳特·阿达巴山·安娜特提莫门·艾拉约阿夫·佩雷格
Owner XIAMEN UNIV
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