Exterior guiding gene T6 concurrently used with RnaseP ribozyme and its uses

An external guidance and combination technology, applied in the fields of molecular biology and medicine, can solve the problems of inability to achieve efficient guidance and cutting, difficulties, etc., and achieve the effect of manual operation and control, improve efficiency, and ensure stability and efficiency.

Inactive Publication Date: 2006-07-26
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, since many mRNAs in the cell are combined with proteins and exist in a highly organized and folded spatial structure, under laboratory conditions where accurate analysis of the secondary structure region of the substrate RNA is not yet possible , it is extremely difficult to select a target site on the substrate RNA that is easy to bind to EGS
Even if you choose a target with high cleavage activity in the M1GS experiment, under the influence of other factors, it may not be able to achieve efficient guided cleavage in the EGS experiment

Method used

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  • Exterior guiding gene T6 concurrently used with RnaseP ribozyme and its uses
  • Exterior guiding gene T6 concurrently used with RnaseP ribozyme and its uses
  • Exterior guiding gene T6 concurrently used with RnaseP ribozyme and its uses

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Sources of plasmids and strains:

[0036] The vector plasmids pGEM3z, pUC19, and the host Escherichia coli JM109 and DH-5a were purchased from Gene Corporation, and they are commonly used vector plasmids and host bacteria in the field.

[0037] The main reagents are shown in Table 1:

[0038] Endonuclease

Dalian Baosheng Company

T4 DNA Ligase

Dalian Baosheng Company

Mung bean sprout nuclease

Dalian Baosheng Company

T7 RNA polymerase

Gene company

Ex Taq polymerase

Dalian Baosheng Company

RNase inhibitor

Dalian Baosheng Company

RNase-free DNase I

Dalian Baosheng Company

rNTP, bovine serum albumin component V

Beijing Dingguo Company

RNaseA

Beijing Dingguo Company

Tris saturated phenol, water saturated phenol

Beijing Dingguo Company

Electrophoresis grade urea

Beijing Dingguo Company

Wide Range DNA Ladder Marker

Dalian Baosheng Company

Nucl...

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Abstract

This invention discloses an external guiding gene and its application, wherein the gene is used combining with the RNase P ribozyme. The related external guiding sequence is based on the RNA, which has the sequence of the SEQ ID NO: 1, and it also includes the external guiding sequence based on the DNA, which has the sequence of the SEQ ID NO: 2, all of the sequences has the special guiding cutting activity that guides RNase P to cut HCMV polyenzyme mRNA flake, and the target point is at 2878-2894 of the UL54 gene. The EGS in this invention is to research and exploit the HCMV infection disease treatment and the gene drugs of related disease.

Description

Technical field [0001] The invention belongs to the field of molecular biology and medicine, and specifically relates to an external guide gene (EGS) combined with RNase P ribozyme and its application in preparing anti-HCMV drugs. Background technique [0002] In the 1980s, Sidney Altman et al. discovered that M1RNA, the RNA component of Escherichia ribonuclease P (RNase P), can independently shear the precursor tRNA (ptRNA). People call RNA with catalytic cleavage activity "ribozyme". The discovery of ribozymes has broadened the horizons of gene therapy. The complementary cutting characteristics and cycling characteristics of ribozymes have obvious advantages in mammalian cell gene therapy. Experimental results in recent years have shown that ribozymes as a tool for gene therapy have broad application prospects and great clinical practical value. [0003] RNase P is a nucleoprotein complex that is widely found in bacteria, yeasts, animals and plants. It is a natural ribozyme pos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/63A61K48/00A61P31/12
Inventor 周天鸿吕静竹李月琴李弘剑
Owner JINAN UNIVERSITY
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