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Oral recombinant yeast and application of target enteric canal DC (dentritic cell) presenting shRNA (short hairpin ribonucleic acid) mediated thereby

A technology of recombinant yeast and intestinal tract, applied in the fields of immunology and genetic engineering, can solve the problems of long time, high cost, complicated operation, etc., and achieve the effect of overcoming the long time.

Inactive Publication Date: 2014-04-02
NORTHWEST A & F UNIV
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Problems solved by technology

Traditional DC cell therapy uses the patient's own monocytes to induce the generation of DCs in vitro, and then loads the corresponding tumor antigens to make DCs loaded with tumor antigens, and then injects these DC cells into the body to stimulate the tumor-killing lymphocytes in the body. Cell proliferation plays a role in tumor monitoring and tumor killing to achieve the goal of eradicating tumors. However, this method is cumbersome, time-consuming, and expensive. Therefore, it is urgent to find a fast, safe and in vivo treatment of DC cells. Way

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  • Oral recombinant yeast and application of target enteric canal DC (dentritic cell) presenting shRNA (short hairpin ribonucleic acid) mediated thereby
  • Oral recombinant yeast and application of target enteric canal DC (dentritic cell) presenting shRNA (short hairpin ribonucleic acid) mediated thereby
  • Oral recombinant yeast and application of target enteric canal DC (dentritic cell) presenting shRNA (short hairpin ribonucleic acid) mediated thereby

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Embodiment Construction

[0029] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0030] 1. Construction of JMB84-hU6 vector

[0031] 1. Using human genomic DNA as a template, use Clone ManagerV7 software to design primers, design primers hU6-F and hU6-R that can specifically amplify the hU6 promoter sequence, and introduce KpnI restriction sites into the upstream primer hU6-F , a SalI restriction site was introduced into the downstream primer hU6-R. The sequence design is shown in Table 1-1:

[0032] Table 1-1 PCR amplification primers of hU6 promoter gene

[0033] Primer name

Primer sequence

hU6-F

5′-TTG GGTACC CCCGAGTCCAACACCCGTGGG-3′

hU6-R

5′-CTA GTC GAC TAGTATATGTGCTGCCGAAGCG-3′

[0034] Note: The underlined part in the hU6-F primer sequence is the KpnI restriction site; the underlined part in the hU6-R primer sequenc...

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Abstract

The invention discloses oral recombinant yeast and an application of target enteric canal DC (dentritic cell) presenting shRNA (short hairpin ribonucleic acid) mediated thereby. Yeast strain is taken as a host cell, and an shRNA interference vector is transfected into the host cell and has element arrangement of a promoter shRNA-miR30. The oral recombinant yeast can pass through the gastrointestinal tract of a live creature, the method for presenting shRNA by the target enteric canal DC can effectively overcome the defect in the prior art during presenting shRNA into the DC, and can be widely used in gene therapy.

Description

technical field [0001] The invention belongs to the technical fields of immunology and genetic engineering, and relates to an oral recombinant yeast and its mediated application of targeting intestinal DC to present shRNA. Background technique [0002] Gene therapy (Gene Therapy) refers to the application of genetic engineering technology to introduce normal genes into patient cells to correct the defects of disease-causing genes and cure diseases. The way to correct it can be to repair the defective gene, or to regulate the expression of the abnormal gene according to the demand, so as to realize the treatment of the defective gene. Commonly used gene therapy is divided into two categories, one is gene correction and gene replacement, that is, gene targeting technology recombines exogenous normal genes in specific parts, so that defective genes can be specifically repaired in situ. The other type is gene enhancement and gene suppression, that is, without knocking out abnor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18A61K48/00A61P35/00C12R1/865
Inventor 张智英张龙邵斯旻徐坤李欣憶徐华荣
Owner NORTHWEST A & F UNIV
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