Application method of long-chain non-coding ribonucleic acid (RNA) gene in preparation of interference inhibitor

A long-chain non-coding, RNA interference technology, applied in the application field of long-chain non-coding RNA gene sequences, can solve the problem of unknown exact mechanism of development and transfer, and achieve far-reaching and popularization effects.

Active Publication Date: 2013-06-19
CENT SOUTH UNIV
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Problems solved by technology

Hepatocellular carcinoma (HCC) is a major type of primary liver cancer. Although new progress has been made in the diagnosis and treatment of hepatocellular carcinoma, there are still many questions about the exact mechanism of its occurrence, development and metastasis. unknown problem

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  • Application method of long-chain non-coding ribonucleic acid (RNA) gene in preparation of interference inhibitor
  • Application method of long-chain non-coding ribonucleic acid (RNA) gene in preparation of interference inhibitor
  • Application method of long-chain non-coding ribonucleic acid (RNA) gene in preparation of interference inhibitor

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Embodiment Construction

[0071] The present invention will be further described below in conjunction with specific embodiments, rather than limiting the present invention.

[0072] 1. New lncRNA gene cloning and analysis

[0073] 1. Materials and methods:

[0074] 1.1 Reagents and kits

[0075] Agarose (agrose) and other commonly used biochemical reagents and gel recovery kits were purchased from Shanghai Huashun Bioengineering Co., Ltd., and 1kbladder DNA molecular weight markers were purchased from Invitrogen; Mini Kit (Qiagen) extraction kit extracts RNA with improved quality, SuperScript TM The III (Invitrogen) kit reverse transcribed RNA into cDNA.

[0076] 1.2 Next Generation Sequencing Technology for RNA Sequencing

[0077] After the total RNA was extracted from human liver cancer tissue samples and normal liver tissue samples, it was purified by column, and the RNA above 200nt was retained;

[0078] Then mRNA is enriched, and mRNA is fragmented by high-temperature heating with divalent ...

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Abstract

The invention relates to an application method of a newly cloned long-chain non-coding ribonucleic acid (RNA) gene, in particular to the application method of the long-chain non-coding RNA method in preparation of an interference inhibitor. The application method of the long-chain non-coding RNA method in preparation of the interference inhibitor comprises designing and combining a short hairpin structure RNA eukaryotic expression vector for closing expression of the long-chain non-coding RNA in an RNA interference mode according to a gene sequence of the long-chain non-coding RNA, and using the short hairpin structure RNA eukaryotic expression vector to successfully restrain the expression of the long-chain non-coding RNA in a liver cancer cell line.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to an application method of a newly cloned long-chain non-coding RNA gene sequence. Background technique [0002] Primary liver cancer is a high-incidence tumor in Asia and some parts of Africa. Its case fatality rate ranks second among malignant tumors in my country, and its five-year survival rate is only about 5%. important topic. Hepatocellular carcinoma (HCC) is a major type of primary liver cancer. Although new progress has been made in the diagnosis and treatment of hepatocellular carcinoma, there are still many questions about the exact mechanism of its occurrence, development and metastasis. Unknown question. [0003] Long non-coding RNA (long non-coding RNA, lncRNA) is a type of non-coding RNA with a transcript length of more than 200 nucleotides. Studies have found that many lncRNAs play an important role in regulating the occurrence and development of ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A61P35/00C12N15/66C12N15/11C12N15/113
Inventor 李桂源熊炜曾朝阳李小玲向波李夏雨黄宏斌廖前进张文玲范松青石磊周鸣马健向娟娟彭淑平周艳宏
Owner CENT SOUTH UNIV
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