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

Interference RNA miniplasmid of hypoxia induction factor

A hypoxia-inducible factor and small interference technology, applied in the field of genetic engineering, can solve the problems of poor RNA chain stability, unfavorable transfection efficiency, easy degradation, etc., and achieve stable and high-efficiency expression

Inactive Publication Date: 2006-09-13
RENJI HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
View PDF0 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, the selection of siRNA fragments is often non-specific, and the RNA chain has poor stability and is easy to degrade, which is not conducive to further improving transfection efficiency.
[0006] Compared with the present invention, the closest prior art is the fragment for HIF-1α designed by Jorg Hanze (Medical School of the Justus-liebig-University) in Germany, but the synthesized one is RNA double-stranded, and the non-stability is higher plasmid

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
  • Interference RNA miniplasmid of hypoxia induction factor
  • Interference RNA miniplasmid of hypoxia induction factor
  • Interference RNA miniplasmid of hypoxia induction factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of siRNA-HIF-1α

[0027] Design of target gene HIF-1α siRNA sequence

[0028] Open the siRNA design tool http: / / www.ambion.com / techlib / misc / siRNA_finder.html, the CDS of human hypoxia-inducible factor 1α (HIF-1α) gene (the CDS (29-2509) of HIF-1α sees SEQID NO: 4) Paste in the input box, select end with TT, and get a series of output sequences of 21 bases (nt) after submission, select 1 to 2 sequences, and confirm that there is no significant similarity with other genes by Blast test , converting the output sequence into two insertion sequences, the sense strand and the antisense strand.

[0029] The selected sequence is as follows:

[0030] The obtained siRNA-upstream sequence is (SEQ ID NO: 1):

[0031] 5′-TGTGAGTTCGCATCTTGATTTCAAGAGAATCAAGATGCGAACTCACATTTTTT-3′

[0032] Among them, TGTGAGTTCGCATCTTGAT is the obtained upstream segment; TTCAAGAGA is a specific loop ring; ATCAAGATGCGAACTCACA is the palindrome structure of the upstream segment. ...

Embodiment 2

[0045] Example 2 Detection of siRNA-HIF-1α

[0046] By electroporation, exogenous siRNA-HIF-1α was transfected into endothelial progenitor cells, and compared with the non-transfected siRNA group as follows.

[0047] 1. Genetic testing

[0048] After 3 days of culture, the cells were transfected with plasmids, incubated in normoxia and hypoxia for 20 hours, and then RT-PCR and β-actin were quantified. The electrophoresis results are shown in Figure 2, in which, lane 1 is a 100bp marker; lanes 2, 4, and 6 are normoxia; lanes 3, 5, and 7 are hypoxia; lanes 2-3 are not transfected cells; lanes 4-5 are pEGFP empty Plasmid-transfected cells; lanes 6-7 are SiRNA-HIF-1α-transfected cells. A is HIF-1α electrophoresis 20 hours after transfection, B is VEGF electrophoresis 20 hours after transfection. The results showed that: HIF1αCDNA exists in both normoxia and hypoxia, and its expression has nothing to do with oxygen concentration. SiRNA interference inhibits HIF1α; VEGF expressio...

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 discloses a low oxygen induction factor low interruption RNA SiRNA-HIF-1 alpha particle. The particle could be high efficiently expressed in cell and could strongly restrict the expression of independent gene in mammal cell.

Description

technical field [0001] The invention belongs to the field of genetic engineering and relates to a genetic engineering plasmid. Furthermore, the invention relates to a hypoxia-inducible factor small interfering RNA plasmid capable of stably and efficiently expressing in cells. Background technique [0002] RNA interference (or called RNA interference, RNA interference, RNAi) refers to the process of degrading the specific mRNA paired with it induced by double-stranded RNA (dsRNA) in cells. In some biological systems, a small number of copies of dsRNA can lead to the degradation of all homologous genes in the cell. This phenomenon is conserved and widespread during evolution, and many genes play important roles in this process. This phenomenon is widespread in nematodes, insects, mammals, plants and fungi. [0003] When the long-chain dsRNA enters the cell, it is recognized by a Dicer nucleolytic enzyme, cut into a small interfering RNA (SiRNA) of 21-25 bases, and the double...

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
IPC IPC(8): C12N15/63C12N15/66
Inventor 王长谦姜萌
Owner RENJI HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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