Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

siRNA capable of targeting human tspan8 gene and application of sirna

A gene and targeting technology, applied in the direction of DNA / RNA fragments, application, gene therapy, etc., can solve problems such as difficult location of nerve damage, poor efficacy of anti-tumor drugs, etc., and achieve the effect of inhibiting proliferation

Inactive Publication Date: 2017-03-01
EAST CHINA UNIV OF SCI & TECH
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For chemotherapy, due to the existence of the blood-brain barrier, common anti-tumor drugs are not effective
For radiation therapy, there are also positioning difficulties and concerns about nerve damage

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
  • siRNA capable of targeting human tspan8 gene and application of sirna
  • siRNA capable of targeting human tspan8 gene and application of sirna
  • siRNA capable of targeting human tspan8 gene and application of sirna

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 TSPAN8 siRNA sequence design and interference plasmid construction

[0043] Search and download the cDNA sequence of TSPAN8 on the ensembl website (http: / / www.ensembl.org / index.html), use the BLOCK-iT RNA Designer software of Life technologies company (http: / / rnaidesigner.lifetechnologies.com / rnaexpress / ) online design of four siRNA sequences complementary to TSPAN8cDNA. Select the siRNA sequence with the highest score, as follows:

[0044] The sequences of siRNA1-4 are as follows:

[0045] siRNA1

[0046] Sense strand: 5'-GCAAUAUGGGUACGAGUAA-3' (SEQ ID NO.7)

[0047] Antisense strand: 5'-UUACUCGUACCCAUAUUGC-3' (SEQ ID NO.8)

[0048] siRNA2

[0049] Sense strand: 5'-GCAAUGACUCUCAAGCAA-3' (SEQ ID NO.1)

[0050] Antisense strand: 5'-CGUUACUGAGAGUUCGUU-3' (SEQ ID NO.2)

[0051] siRNA3

[0052] Sense strand: 5'-GGAAGCCAUAAUUGUGUUU-3' (SEQ ID NO.9)

[0053] Antisense strand: 5'-AAACACAAUUAUGGCUUCC-3' (SEQ ID NO.10)

[0054] siRNA4

[0055] Sense strand:...

Embodiment 2

[0077] Example 2: Interfering with the packaging of lentiviral particles

[0078] 5% CO at 37°C 2 HEK 293T cells (hereinafter referred to as 293T, purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences) were cultured in a cell incubator. Produced by the company) culture medium. Subculture 293T cells in a petri dish with a diameter of 10 cm. When the cells grow to a confluence of about 50%, culture them with serum-free medium for 4 hours.

[0079] Operate according to the instructions of Lipofectamine 2000 (produced by U.S. Invitrogen Corporation), prepare the pLKD-CMV-GFP-U6-shRNA plasmid (or empty vector plasmid without shRNA) obtained above 22.5g, 7.9g virus coat plasmid psPAX2 (Shanghai New York, China) En Biotechnology Co., Ltd.), 14.6 g of the transfection mixture of the packaging plasmid pMD2.G (provided by Shanghai Nuen Biotechnology Co., Ltd., China).

[0080] Add the mixture to starved cultured cells for transfection...

Embodiment 3

[0082] Example 3: Verification of interference efficiency at the protein level

[0083] Protein sample preparation:

[0084]The cells were cultured in a 6 cm diameter petri dish to about 30% confluence, the control lentivirus infection particles and the virus infection particles interfering with TSPAN8 were added respectively (the multiplicity of infection was 10), and the cells were continued to be cultured for 4 days. The experiment was divided into 5 groups, namely: negative control virus infection group (Control), Y2037 virus infection group, Y2038 virus infection group, Y2039 virus infection group and Y2040 virus infection group. When collecting the cells, transfer the medium to an empty 15ml centrifuge tube, rinse the cells twice with PBS, each time with 3ml of PBS, and then digest the cells with 2ml of 0.25% trypsin. The medium was poured back into the Petri dish. Transfer the digested cells along with the medium to a 15ml centrifuge tube, and centrifuge at 1200 rpm f...

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 relates to the fields of biological medicines and gene therapy; the invention provides siRNA or shRNA capable of specifically inhibiting the expression of the TSPAN8 gene, and provides an application of the siRNA or shRNA in preparing a TSPAN8 gene expression inhibitor as well as an application in preparing medicines for preventing or treating gliomas.

Description

technical field [0001] The invention relates to the fields of biomedicine and gene therapy, in particular to an siRNA targeting human TSPAN8 gene and its application in the preparation of antitumor drugs. Background technique [0002] The full English name of TSPAN8 is tetraspanin 8 (tetraspanin 8), also known as CO-029, TM4SF3, located on chromosome 12 (Genebank Accession: NM_004616). TSPAN8 is one of the important members of the four-span membrane superfamily. TSPAN8 affects phagocytic glycoprotein 1 (CD44) (Guo Q.ect, Tetrasoanin CO-029 inhibit colorectal cancer cell movement by deregulationg cell-matrix and cell-cell adhensions, PloS One.2012; 7(6)e:38464) , or affect local focal adhesion kinase (FocalAdhesion Kinase, FAK), or affect paxilin (Paxilin) ​​(Shijing Yue et al., Tspan8and CD151promote metastasis by distinct mechanisms. European Journal of Cancer (2013) 49, 2934–2948), and then affect The expression of laminin-binding intergrin α3β1 and fibronectin-intergrin...

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 Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/63C12N15/867C12N15/861A61K48/00A61P35/00
Inventor 祖勇郑静杨艳红朱志川李奎刘霁纬
Owner EAST CHINA UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com