Method for screening natural antisense transcripts related to human tumor

An antisense transcript, natural technology, applied in the field of screening human tumor-related natural antisense transcripts, can solve the problem of unreliable results and achieve the effect of effective screening

Inactive Publication Date: 2015-02-04
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of these studies use the method of bioinformatics prediction, which has the advantages of convenience and speed, but the results obtained are not reliable enough, and the differentially expressed NAT molecules cannot be systematically screened in specific cells or tissues.

Method used

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  • Method for screening natural antisense transcripts related to human tumor
  • Method for screening natural antisense transcripts related to human tumor
  • Method for screening natural antisense transcripts related to human tumor

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Embodiment Construction

[0034] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0035] Such as figure 1 As shown, the method for screening human tumor-associated natural antisense transcripts provided by the invention comprises the following steps:

[0036] 1. Collection of cell culture / tissue specimen materials

[0037] Cultivate the target cells according to the optimized culture conditions of various cells; if using tissue sample materials, the tissue sample materials should be stored in liquid nitrogen immediately after operation.

[0038] 2. Extraction of total RNA

[0039] The total RNA was extracted with the total RNA extraction reagent Trizol, and the operation step...

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Abstract

The invention provides a method for screening natural antisense transcripts related to the human tumor. The method comprises the following steps: extracting cancer cell / normal cell total RNA (ribonucleic acid); performing DNase I enzyme cutting and RNase I enzyme cutting; performing reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), performing double-chain synthesis, and adding linkers; performing suppression subtractive hybridization, cloning the polymerase chain reaction (PCR) product to establish a library, and performing sequencing analysis; and further performing experimental verification to candidate molecules and the like. After the method is applied and verified in normal cells and tumor cells, the method is proven to screen the natural antisense transcripts related to human tumor systematacially and rapidly.

Description

technical field [0001] The invention relates to the fields of biotechnology and biomedicine, in particular, the invention relates to a method for screening human tumor-associated natural antisense transcripts. Background technique [0002] Natural antisense transcripts (natural antisense transcripts, hereinafter referred to as NAT) usually refer to antisense RNAs (antisense RNAs) produced in organisms under natural conditions, and they form a double pair with their corresponding complementary RNAs (complementary RNAs) through complementary base pairing. The natural sense-antisense transcript (natural sense-antisense transcript, hereinafter referred to as NSAT) of the chain can cause the degradation or translation inhibition of the target gene to regulate the expression of the target gene. NAT is divided into two types according to its mode of action: most NATs originate from the same genomic locus as the positive-sense transcript, and are encoded by the opposite strand encod...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/10C12Q1/68
Inventor 杨朝辉何凤田张立
Owner ARMY MEDICAL UNIV
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