Long non-coding RNA sequence relevant to human melanoma cells and application thereof

A technology for melanoma cells and melanoma, applied in the field of long non-coding RNA related to human melanoma cells and its application

Inactive Publication Date: 2011-04-13
SICHUAN UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These research results indicate that lncRNA plays an important role in cell proliferation, differentiation and carcinogenesis, and the discovery of tumor-associated lncRNA may have a significant impact on tumor diagnosis and gene therapy

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Long non-coding RNA sequence relevant to human melanoma cells and application thereof
  • Long non-coding RNA sequence relevant to human melanoma cells and application thereof
  • Long non-coding RNA sequence relevant to human melanoma cells and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Trizol reagent was purchased from Invitrogen Company of the United States; MMLV reverse transcriptase, DNA polymerase, pfu enzyme, NdeI enzyme, XhoI enzyme and RNase inhibitor were purchased from Lithuanian Fermentas Company; T7 RNA polymerase was purchased from NEB Company of the United States; random primers were purchased from the United States Promega Company; TALON metal ion resin was purchased from BDClontech Company of the United States; PCR primers were synthesized by Invitrogen Company.

[0025] 2. Bacterial strains, cell lines and vectors

[0026] Human malignant melanoma cells yusac were purchased from American Type Culture Collection (ATCC); E.coliBL2 strain was purchased from Tianjin Tianli Technology Co., Ltd.; PCR-blunt-TOPO vector was purchased from Invitrogen, USA; pET-28a(+) vector Purchased from Novagen, Germany.

[0027] 3. Experimental method

[0028] 3.1 Construction of human malignant melanoma cell yusac total cDNA library

[0029] 3.1.1 Extrac...

Embodiment 2

[0109] The conventional reagents, bacterial strains, cell strains and vectors are all the same as in Example 1.

[0110] QuantiTectSYBRGreenPCR kit was purchased from Qiagen, Germany.

[0111] 2. Experimental method

[0112] 2.1.1 Synthesized total cDNA

[0113] Using human melanoma tissue and corresponding paracancerous tissue as materials, total RNA was extracted from melanoma tissue and corresponding paracancerous tissue with Trizol reagent (Invitrogen) (the method is the same as 3.1.1 in Example 1). Take 1 μg of the total RNA of the melanoma tissue and the total RNA of the corresponding paracancerous tissue, and carry out reverse transcription reaction with random primers (the method is the same as 3.1.2 in Example 1) to obtain the total cDNA of the melanoma tissue and the corresponding paracancerous tissue, and synthesize The total cDNA of the melanoma tissue and the corresponding paracancerous tissue were the templates for the quantitative PCR reaction.

[0114] 2.1.2...

Embodiment 3

[0120] The conventional reagents, bacterial strains, cell strains and carriers are the same as in Example 1.

[0121] pcDNA-3.1 eukaryotic expression vector, lipofectamine2000 transfection reagent, and G418 antibiotics were all purchased from Invitrogen Company of the United States; DMEM medium and fetal bovine serum were purchased from Gibico Company of the United States; nitroblue tetrazolium (NBT) was purchased from Sigma Company of the United States. company.

[0122] Perfection4990 flatbed scanner was purchased from Epson Corporation of Japan.

[0123] BALB / c nude mice, SPF (Specific Pathogen-free, specific pathogen-free) grade, were purchased from Shanghai Slack Experimental Animal Co., Ltd.

[0124] 2. Experimental method

[0125] 2.1 Construction of monoclonal cell lines stably and highly expressing lncRNA-HN3

[0126] 2.1.1 Construction of eukaryotic recombinant plasmid pcDNA-lncRNA-HN3 of lncRNA-HN3

[0127] The DNA sequence (obtained by 3.3 and 3.4 of Example 1)...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a long non-coding RNA (long non-coding RNA, 1ncRNA)-incRNA-HN3 sequence relevant to human melanoma cells, and an RNA interference (RNA interference, RNAi) target spot sequence, a short-hairpin RNA (short-hairpin RNA, shRNA) sequence, an shRNA coded small molecule interference RNA (small interference RNA, siRNA), a coding shRNA coding strand and an anti-sense strand DNA sequence, coding shRNA recombinant plasmids of the long non-coding RNA sequence, and the invention relates to the applications of the long non-coding RNA sequence for preparing reagents for diagnosing melanoma diseases and drugs for treating the melanoma diseases.

Description

technical field [0001] The invention belongs to the professional field of biomedicine, and relates to a novel lncRNA belonging to a human lncRNA, its RNA interference (RNA interference, RNAi) target sequence, a short-hairpin RNA (short-hairpin RNA, shRNA) sequence, and a small RNA encoded by shRNA. Molecular interference RNA (small interference RNA, siRNA), sense and antisense strand DNA encoding shRNA, eukaryotic recombinant plasmid encoding shRNA, and their use in the preparation of reagents for diagnosing melanoma diseases and the preparation of drugs for treating melanoma diseases. Background technique [0002] There are about 20,000 to 30,000 genes that encode proteins in the human body, accounting for only 2% of the human genome, and the remaining 98% of genomic DNA that does not encode proteins was originally considered to be nonfunctional and garbage in organisms, often called "Junk DNA". However, current research shows that most of these junk DNA can be transcribe...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/79C12Q1/68A61K48/00A61P35/00C12N15/113
Inventor 宋旭李灵俸婷婷连莹莹张广丰
Owner SICHUAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap