Surefection Technology for High-Throughput Primary Cell Transfection

高通量原代细胞转染的 Surefection 技术

• 批准号: 7053098
• 负责人: YONG HUANG
• 金额: $ 19.97万
• 依托单位: RATIONAL BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7053098
• 关键词: electroporation; high throughput technology; human; transfection

DESCRIPTION (provided by applicant): Human primary cells are cells directly derived from tissue samples from patients or healthy donors. They are the truest in vitro models of human physiology and disease, therefore are invaluable in medical discovery and development. Use of human primary cells as physiology- or disease-relevant models provides significant advantages in research and development for better understanding and treatment of various human diseases, including cancer, cardiovascular and inflammatory diseases. Unfortunately, utility of precious primary cells is largely hindered by the lack of means for effective transfection with minimal side-effects, particularly in high- throughput mode. To address this unmet need, we propose the development of a SureFection technology that combines two emerging advancements in the field of cell transfection, i.e., controlled in situ electroporation and DMA nuclear delivery, to effective transfect primary cells free of biochemical side-effects. This strategy will be implemented in novel miniature devices that can be readily multiplexed to conform to 96-well slide/1536-well microplate uHT footprints, permitting massive scale in vivo gene funtion analysis, as well as drug screening and testing using primary cells. In Phase I, we will demonstrate the applicability of our technology in three primary cell systems that are important models for study of breast and prostate cancer, angiogenesis and cardiovascular disease. Our long-term goal is to combine this enabling technology with our novel functional assay methods to provide primary cell based high-throughput solutions for target ID&validation, compound screening and in vitro drug testing/profiling, as part of our strategies to facilitate faster, cheaper and safer small molecule drug development. The proposed research and development will benefit public health by forwarding our understanding of the roles of human genes in physiology and disease, and accelerating therapeutic development to fight with cancer, cardiovascular and other diseases.

描述（由申请方提供）：人原代细胞是直接来源于患者或健康供体组织样本的细胞。它们是人体生理学和疾病的最真实的体外模型，因此在医学发现和发展中是无价的。使用人原代细胞作为生理学或疾病相关模型在研究和开发中提供了显著的优势，以更好地理解和治疗各种人类疾病，包括癌症、心血管和炎性疾病。不幸的是，珍贵的原代细胞的实用性在很大程度上受到缺乏具有最小副作用的有效转染手段的阻碍，特别是在高通量模式下。为了解决这一未满足的需求，我们提出开发一种SureFection技术，该技术结合了细胞转染领域的两个新兴进展，即，控制原位电穿孔和DMA核递送，以有效地去除原代细胞而没有生化副作用。该策略将在新型微型装置中实施，该装置可以容易地多路复用以符合96孔载玻片/1536孔微孔板uHT足迹，从而允许大规模体内基因功能分析以及使用原代细胞的药物筛选和测试。在第一阶段，我们将证明我们的技术在三种原代细胞系统中的适用性，这些系统是研究乳腺癌和前列腺癌、血管生成和心血管疾病的重要模型。我们的长期目标是将这种使能技术与我们的新型功能测定方法联合收割机相结合，为靶点识别和验证、化合物筛选和体外药物测试/分析提供基于原代细胞的高通量解决方案，作为我们促进更快、更便宜和更安全的小分子药物开发战略的一部分。拟议的研究和开发将通过促进我们对人类基因在生理学和疾病中的作用的理解，并加速治疗癌症，心血管和其他疾病的治疗发展，使公众健康受益。