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Application of TRIM28 gene and its expression product

A technology for expressing products and gene expression, which is applied in the field of gene application and can solve the problems of unsatisfactory clinical efficacy of biological therapy

Inactive Publication Date: 2012-09-26
CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the clinical efficacy of many biological treatments is still not very satisfactory, and most of them are still in the stage of experimental research, and the relevant key theories and experimental techniques still need further research and development.

Method used

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  • Application of TRIM28 gene and its expression product
  • Application of TRIM28 gene and its expression product
  • Application of TRIM28 gene and its expression product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Kinome-wide RNAi screening of essential kinase genes for liver cancer cell growth

[0034] As a simple, rapid, specific, efficient, economical, and predictable technology, RNAi has obvious advantages. It is superior to antisense nucleic acid technology, simpler than gene knockout, and has a good application prospect. Specifically, it can be used in the following aspects: gene function analysis; new ways to study signal transduction pathways; research and development of new drugs; gene therapy. RNAi only inhibits the disease-causing gene without affecting the function of the normal allele. It has high selectivity and specificity, and can reduce the side effects caused by non-specific effects. Tumor occurrence is the result of multi-gene interaction gene network regulation. The blockade of a single oncogene induced by traditional techniques cannot completely inhibit or reverse the growth of tumors, but RNAi can take advantage of the homology of multiple genes in the same ...

Embodiment 2

[0037] Example 2 shRNAs interfere with the TRIM28 gene to inhibit the growth of liver cancer cells

[0038] 48 hours after the constructed plasmid pSUPER was transfected into liver cancer cells, the cell line RNA was extracted, and the effect of RNA interference was identified by quantitative PCR. For transfection, the cells were re-seeded in a 10cm large plate for culture, and neomycin G418 was added to the culture medium for selection and culture for 4 to 6 weeks. The culture medium was removed, washed twice with 1×PBS, stained with crystal violet, and photographed for counting. The experimental results showed that quantitative PCR detection of shRNAs effectively silenced the expression of the TRIM28 gene, and significantly inhibited the ability of cell colony formation.

Embodiment 3

[0039] Example 3 Immunoassay

[0040] 1. Acquisition of antigenic protein

[0041] (1) Expression by genetic engineering: the cDNA sequence of the human TRIM28 gene can be obtained from the Genebank database, the coding frame can be obtained by PCR amplification, inserted into a prokaryotic or eukaryotic expression vector, and the TRIM28 protein can be expressed, and the product can be expressed according to genetic engineering Purification system for protein purification.

[0042] 2. Antibody Preparation

[0043] Antibodies can be prepared in several ways:

[0044] (1) Cell fusion method: immunize animals (including rabbits, goats, etc.) with the TRIM28 protein prepared above, obtain spleen cells, fuse with myeloma cells, and prepare monoclonal antibodies according to conventional monoclonal antibody preparation techniques.

[0045] (2) Using the phage surface display library, cloning the spleen IgG variable region of the immunized animal and expressing it into a genetical...

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Abstract

The invention discloses application of a TRIM28 gene and its expression product in preparing liver cancer treatment products. The TRIM28 gene and its expression product can be taken as a target gene for preparing medicaments treating liver cancer, thus providing a new approach for liver cancer treatment.

Description

technical field [0001] The present invention relates to the application of a gene, in particular to the application of a TRIM28 gene and its expression product. Background technique [0002] Protein kinase is encoded by the largest class of genes in the human genome. Its function is to regulate the activity of various proteins, signal transduction pathways and cell biological processes by transferring phosphate groups to substrate proteins. Human genome Encoding 518 protein kinases among more than 57 kinase families, approximately 55% of known proto-oncogenes encode protein kinases, and most of the remaining proto-oncogenes encode specific proteins that activate protein kinases or are phosphorylated by kinases. All protein kinases are involved in cell signaling pathways, involved in the pathogenesis of cardiovascular diseases, diabetes, infectious diseases, arthritis and other immune disorders, nervous system such as Alzheimer's disease, Alzheimer's disease AD, etc., have be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/48G01N33/573A61K45/00A61K38/00A61K39/395A61K48/00A61P35/00C12N15/113
Inventor 韩泽广邓庆李坤雨
Owner CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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