Retrovectors for Intron-Enhanced Protein Expression

用于内含子增强蛋白质表达的逆向载体

• 批准号: 6991099
• 负责人: MICHAEL IMBODEN
• 金额: $ 8.97万
• 依托单位: IOGENETICS, LLC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6991099
• 关键词: CHO cells; Hepadnaviridae; Retroviridae; biological transport; bovine leukemia virus; complementary DNA; gene delivery system; immunoglobulin genes; introns; monoclonal antibody; polymerase chain reaction; posttranslational modifications; technology /technique development; transfection /expression vector; virus RNA

DESCRIPTION (provided by applicant): Retrovectors are widely used for gene transfer in gene therapy, tissue culture and transgenic animals for research or commercial applications.. A long recognized limitation of current retrovectors is that introns are spliced out as retrovector particles are made in packaging cells. Introns may regulate the expression of the genes and convey tissue specificity. The presence of introns boosts expression as a consequence of active splicing. This proposal tests the design of retrovectors which preserve introns by utilizing elements of viral RNA export mechanisms naturally occurring in the complex retroviruses. A packaging cell expressing an RNA export enabling protein (Rex) will be used to produce vectors whose constructs contain the corresponding Rex responsive element (RxRE). In combination this should allow the making of retroviral vectors which contain introns. These vectors subsequently allow the transfer of introncontaining constructs into the final host cell. Relevance: Previously considered "junk DMA", introns are now understood to exert significant effects on the adjacent coding regions of genes. By enabling the transfer of intron-containing genes we anticipate (a) providing a means to make gene therapy vectors with greater predictability of expression and tissue specificity; (b) providing a boost to protein expression in cell culture bioreactor systems and transgenic animals and (c) providing a useful research tool to enhance understanding of RNA biology.

描述（由申请人提供）：