Supercharge Your Innovation With Domain-Expert AI Agents!

Tissue-specific double gene silencing RNAi expression vector having self-splicing locus

A tissue-specific, expression vector technology, applied in the medical field, can solve problems such as non-tissue-specific effects, limit gene silencing, etc., and achieve the effect of simple construction method and easy operation.

Inactive Publication Date: 2013-09-11
ZHENGZHOU UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the above situation, in order to overcome the defects of the prior art, the object of the present invention is to provide a tissue-specific double-gene silencing RNAi expression vector comprising a self-cleavage site, which can effectively solve the problem of RNAi expression vector transfection of mammalian cells. Limitations when simultaneously inhibiting two genes and problems with non-tissue-specific effects during gene silencing

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
  • Tissue-specific double gene silencing RNAi expression vector having self-splicing locus
  • Tissue-specific double gene silencing RNAi expression vector having self-splicing locus
  • Tissue-specific double gene silencing RNAi expression vector having self-splicing locus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Insertion of the first oligonucleotide with self-cleaving ribozyme sequence:

[0041] In order to facilitate the insertion of the initial vector pEGFP-C1, a Nhe Ⅰ restriction site (GCTAGC) was added to the 5'end of the first cis-acting cis-acting hammerhead ribozyme sequence (SEQ ID NO:1), at the 3'end Add the BglⅡ restriction site (AGATCT), artificially synthesize it, and obtain the first cis-acting non-hook hammerhead shark ribozyme sequence oligonucleotide double-stranded with restriction enzymes Nhe Ⅰ and Bgl Ⅱ , Insert the Nhe Ⅰ and Bgl Ⅱ restriction sites of the original vector pEGFP-C1 to construct the vector pRib (SEQ ID NO: 7);

[0042] (2) Insertion of the second oligonucleotide with self-cleaving ribozyme sequence:

[0043] To facilitate the insertion into the vector pRib, add HindⅢ restriction site (AAGCTT) to the 5'end of the second cis-acting Hammerhead ribozyme sequence (SEQ ID NO:1), and add EcoR Ⅰ restriction to the 3'end Site (GAATTC), artificially synt...

Embodiment 2

[0056] (1) Insertion of the first oligonucleotide with self-cleaving ribozyme sequence:

[0057] In order to facilitate the insertion of the initial vector pEGFP-C1, a Nhe Ⅰ restriction site (GCTAGC) was added to the 5'end of the first cis-acting cis-acting hammerhead ribozyme sequence (SEQ ID NO:1), at the 3'end Add the BglⅡ restriction site (AGATCT), artificially synthesize it, and obtain the first cis-acting non-hook hammerhead shark ribozyme sequence oligonucleotide double-stranded with restriction enzymes Nhe Ⅰ and Bgl Ⅱ , Insert the Nhe Ⅰ and Bgl Ⅱ restriction sites of the original vector pEGFP-C1 to construct the vector pRib (SEQ ID NO: 7);

[0058] (2) Insertion of the second oligonucleotide with self-cleaving ribozyme sequence:

[0059] To facilitate the insertion into the vector pRib, add HindⅢ restriction site (AAGCTT) to the 5'end of the second cis-acting Hammerhead ribozyme sequence (SEQ ID NO:1), and add EcoR Ⅰ restriction to the 3'end Site (GAATTC), artificially synt...

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 a tissue-specific double gene silencing RNAi expression vector having a self-splicing locus and solves the problem of limitation on the RNAi expression vector inhibiting two genes at the same time during transfection of a mammalian cell and the problem of non-tissue-specific action in a gene silencing process. The vector has a tissue-specific double gene silencing shRNA expression cassette. The tissue-specific double gene silencing shRNA expression cassette comprises multiple cloning sites to be inserted into a tissue-specific promoter, two ribozyme sequences containing self-splicing loci, two multiple cloning sites to be inserted into an shRNA template sequence and a transcription terminator sequence.

Description

Technical field [0001] The present invention relates to the medical field, in particular to a tissue-specific double-gene silencing RNAi expression vector containing self-cutting sites. Background technique [0002] RNA interference (RNAi) refers to small RNA double-stranded (short interfering RNA, siRNA) with a specific sequence after being transported to cells by inducing degradation of homologous messenger RNA (mRNA), resulting in transcription of specific genes The phenomenon of post-gene silencing (Fire et al, Nature1998391:806). siRNA is a ribonucleic acid (RNA) double-stranded molecule with a two-base redundant 3'hydroxyl end. This molecule can bind to the silencing complex and cleave the target gene mRNA. Cleavage usually occurs in the middle of the homologous part of the siRNA, 10 bases from the 5'end of the siRNA antisense strand. siRNA is not only easily degraded by RNases that are widely present in the environment, but also has the disadvantages of poor stability, s...

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): C12N15/63C12N15/66C12N15/86
Inventor 任雪玲张振中柴丽娜宋雨张云
Owner ZHENGZHOU UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com