Blood vessel growth hormone gene recombined gland relative vrius carrier and its preparation process and application thereof
An angiogenin and gene recombination technology, applied in the field of genetic medicine, can solve the problems of adenovirus contamination, application, obstacles, etc., and achieve the effects of low equipment requirements, immune response avoidance, and wide application range
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0030] Example 1: Preparation (packaging) of rAAV-ANG of the present invention:
[0031] According to the ANG gene sequence and the structure of plasmid pAAV-MCS, design primers for the angioglossin gene with restriction enzymes BamHI and EcoRI restriction sites, and amplify the angiogenin ring deoxyribonucleic acid fragment (398bp) by polymerase chain reaction ), connected to the plasmid pAAV-MCS to construct the plasmid pAAV-ANG, restriction enzyme digestion and sequencing to identify the insert direction and sequence are correct (seefigure 1 ). Aspirate 10μg each of plasmids pAAV-ANG, pAAV-RC and pHelper, and use calcium phosphate co-precipitation method to transfect 293 cells to construct recombinant angiogenin gene adeno-associated virus vector (rAAV-ANG); at the same time establish a positive control, namely pAAV-LacZ 10μg each of pAAV-RC and pHelper was co-transfected into 293 cells to construct a recombinant reporter gene LacZ virus vector (rAAV-LacZ). The recombinant vir...
Embodiment 2
[0032] Example 2: Recombinant virus rAAV-ANG is used to study the capillary density, left ventricular remodeling and cardiac function of normal rat myocardial tissue:
[0033] Rats were given general anesthesia by intraperitoneal injection of chloral hydrate (400mg / kg), taken supine position, cut the trachea, connected to the animal ventilator, opened the chest cavity, and exposed the heart. Pipette 120μL of rAAV-ANG (about 1.2×10 9 IU) was injected at four points on the anterior wall of the left ventricle, 30 μL at each site. Close the chest cavity and remove the ventilator. After the rats were awake, they were fed with ordinary feed in a clean environment for four weeks. The results showed that rAAV-ANG efficiently transfected normal rat myocardial tissue and significantly up-regulated the expression of angiogenin protein in normal rat myocardial tissue (see figure 2 ), increase the capillary density in the myocardial tissue (see image 3 ). But it did not affect the ventricular...
Embodiment 3
[0034] Example 3: Research on the use of recombinant virus rAAV-ANG for gene therapy in rats with myocardial infarction:
[0035] Rats were given general anesthesia by intraperitoneal injection of chloral hydrate (400mg / kg), taken supine position, cut the trachea, connected to the animal ventilator, opened the chest cavity, and exposed the heart. Open the pericardial wall, quickly squeeze the heart out of the chest cavity, and ligate the left anterior descending coronary artery with an 8-0 silk thread between the left atrial appendage and the right ventricular outflow tract, resulting in acute myocardial infarction. Then quickly put the heart back into the chest cavity. Pipette 120μL of rAAV-ANG (about 1.2×10 9 IU) Intramyocardial injection was performed at four points on the edge of the infarcted area of the left ventricular anterior wall. Close the chest cavity and remove the ventilator. After the rats are awake, they are fed with ordinary feed in a clean environment. Four wee...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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