Gene Vector Core
基因载体核心
基本信息
- 批准号:7488131
- 负责人:
- 金额:$ 21.18万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2008
- 资助国家:美国
- 起止时间:2008-05-01 至 2013-03-31
- 项目状态:已结题
- 来源:
- 关键词:AdenovirusesBiologicalCardiacCardiac MyocytesCell LineCentrifugationCytomegalovirusDependovirusDevelopmentDialysis procedureEnhancersFacility Construction Funding CategoryGenesHela CellsIn VitroMicroRNAsMolecularNeonatalParticipantPhysical DialysisPlasmidsProductionRattusRecommendationSchemeSerotypingServicesSubfamily lentivirinaeSucroseTestingTitrationsTransgenesViralViral VectorVirusWestern Blottingcesium chloridedesigndesign and constructionmembernovel therapeuticsprogramspromotertherapeutic genetransduction efficiencytransgene expressionvector
项目摘要
Gene Vector and Production Core
1. Core Function:
This viral vector core function is to provide each of the four projects two major services:
(1) Design and construction of transgenes for packaging within viral vectors;
(2) Production, purification and testing of those viral vector types.
Design and construction of packaging plasmids include:
A) Each viral vector has specific design requirements including packaging capacity, serotype and pseudotype
recommendations, these will be examined prior to construction of each new packaging plasmid.
B) The construction of each packaging plasmid will be confirmed by restriction digests and sequencing, and
then appropriately tested for transgene expression.
Production, purification and testing include:
A) Production of:
AAV serotypes 1 -9
Adenovirus type 2
VSVG pseudotyped Lentivirus
B) Purification will utilize cesium chloride gradient centrifugation for AAV and Adenovirus and sucrose cushion
gradients for Lentivirus production, check post production purification schemes.
C) Testing of each virus that will include:
i) Titration of the viruses after dialysis prior to delivery to project leaders.
ii) Testing of the in vitro transduction efficiency of viruses in HeLa cells for CMV or other strong
promoters, or primary neonatal rat cardiomyocytes for restricted (cardiac) promoters.
iii) Western blot analysis will be used to establish expression of viral transgenes prior to delivery to
project leaders.
This core will manufacture and purify AAV serotypes 1-9, Adenovirus type 2 and Lentivirus. Specifically,
we will develop and maintain cell lines for large scale (100s of 15-cm plates) production of specific transgenes
as dictated by PPG participant needs. We will provide the molecular biological support related to sub-cloning
of novel therapeutic genes, inhibitory and micro RNAs, as well as enhancer-promoter configurations for viral
development as needed by the Project Leaders. The main functions of Core D are; 1) Production of Adenoand
Adeno-associated viruses, 2) Purification of these viruses, and 3) Testing viruses prior to transferring to
members of the Scientific Program. An additional function will be the production of VSVG pseudotyped
lentivirus, as needed by members of the Scientific Program.
基因载体与生产核心
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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JOSEPH E RABINOWITZ其他文献
JOSEPH E RABINOWITZ的其他文献
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{{ truncateString('JOSEPH E RABINOWITZ', 18)}}的其他基金
Viral Gene Expression to Therapeutic Levels in Models of Heart Failure
心力衰竭模型中病毒基因表达达到治疗水平
- 批准号:
8066612 - 财政年份:2008
- 资助金额:
$ 21.18万 - 项目类别:
Viral Gene Expression to Therapeutic Levels in Models of Heart Failure
心力衰竭模型中病毒基因表达达到治疗水平
- 批准号:
8274859 - 财政年份:2008
- 资助金额:
$ 21.18万 - 项目类别:
Viral Gene Expression to Therapeutic Levels in Models of Heart Failure
心力衰竭模型中病毒基因表达达到治疗水平
- 批准号:
7841837 - 财政年份:2008
- 资助金额:
$ 21.18万 - 项目类别:
Viral Gene Expression to Therapeutic Levels in Animal Models of Heart Failure
心力衰竭动物模型中病毒基因表达达到治疗水平
- 批准号:
7665572 - 财政年份:2008
- 资助金额:
$ 21.18万 - 项目类别:
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