Novel Micro/nanofluidic Electroporation Devices for DNA&Oligonucleotide Delivery

新型 DNA 微/纳流体电穿孔装置

基本信息

批准号：
7498973
负责人：
Ly James Lee
金额：
$ 20.62万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7498973
关键词：
Academia Acceleration Acids Area Biological Models Cell Count Cell Line Cell Survival Cell membrane Cells Clinical Clinical Trials Condition Cultured Cells DNA Devices Digit structure Electroporation Encapsulated Evaluation Gene Delivery Gene Transfer Genes Government Growth Factor In Vitro Localized Marketing Melanoma Cell Membrane Methods MicroRNAs Microcapsules drug delivery system Microfluidics Molecular Probes Motion Mus Nuclear Nucleic Acids Oligonucleotides Operative Surgical Procedures Patients Performance Persons Pharmacologic Substance Pharyngeal structure Physiologic pulse Polymers Population Process Provider Pulse taking Purpose Range Reporter Genes Research Research Proposals Small Interfering RNA Solutions Stem cells System T-Lymphocyte Techniques Technology Testing Therapeutic Time Tissues Transfection Viral Work base blastomere structure cancer cell cancer stem cell clinical application design and construction desire electric field embryonic stem cell experience in vivo knock-down leukemia nanofluidic novel plasmid DNA tool transcription factor two-dimensional uptake voltage

项目摘要

DESCRIPTION (provided by applicant): Electroporation is a widely used transfection method to introduce nuclear acids and other molecular probes into cells or tissues. Recent progress has made this method applicable to a wide range of cells and biomolecules, for in vitro transfection testing and for ex vivo and even in vivo clinical trials. The current worldwide market for electroporation is about $25 million and is expected to grow rapidly with double-digit gains over the next five years, stimulated by academia, government and pharmaceutical and biotech companies. The current commercial electroporation devices and kits (Amaxa Nucleofector technology in particular) are convenient, easy to use, and beneficial for many hard-to-transfect cell lines when non-viral gene transfer is required. However, a major drawback is that each new cell system (such as lab-specific clones or a patient's primary cells) often requires a time consuming and expensive trial-and-error search process to identify proper electroporation conditions and/or solution composition that will achieve the desired transfection level with high enough cell viability. Each cell system must be optimized individually using a finely-tuned, high electric voltage and non-uniform electric field. It would be very valuable if electroporation could be carried out in a mild, uniform electric field that is effective with various cell lines and primary cells. In this way, the need to determine new settings for each cell system can be eliminated or highly simplified. For clinical applications, there is a need to transfect a large number of cells (e.g. >109 cells) with a high transfection efficiency and cell viability. This is difficult to achieve in the current commercial electroporation device where the recommended cell population is around 105 106. We plan to design, construct and characterize a batch-type Nanonozzle Sandwich Electroporation (NSE) device and a continuous-type Converging Flow Electroporation (CFE) device. These two novel devices will then be used to investigate in vitro transfection efficiency and uptake of selected oligonucleotides and genes for cancer and stem cell applications. Their performance will be compared with the best commercial electroporation systems. Several novel electroporation devices will be developed for better in vitro and ex vivo transfection of therapeutic materials. In this study, the uptake of oligonucleotides and plasmid DNA into cancer cells and stem cells will be investigated. The proposed devices have great potential to transfect oligonucleotides and plasmid DNA into a large number of cells for clinical applications.

描述（由申请人提供）：电穿孔是一种广泛使用的转染方法，用于将核酸和其他分子探针引入细胞或组织。最近的进展使得这种方法适用于广泛的细胞和生物分子，用于体外转染测试，外体内甚至体内临床试验。当前的全球电穿孔市场约为2500万美元，预计在未来五年内，学术界，政府和制药和生物技术公司刺激了未来五年的两位数增长。当需要非病毒基因转移时，当前的商用电穿孔设备和试剂盒（尤其是Amaxa核对象技术）方便，易于使用，并且对许多难以转移的细胞系有益。但是，一个主要的缺点是，每个新细胞系统（例如特定于实验室的克隆或患者的原始细胞）通常需要耗时且昂贵的试验搜索过程，以识别适当的电穿孔条件和/或溶液组合物，以实现具有足够高的单元格性的所需转染水平。必须使用精细调整的高电压和不均匀的电场来单独优化每个电池系统。如果可以在各种细胞系和主要细胞有效的轻度，均匀的电场中进行电穿孔，那将是非常有价值的。这样，可以消除或高度简化每个单元系统的新设置的需求。对于临床应用，需要具有高转染效率和细胞活力的大量细胞（例如> 109个细胞）。在当前推荐的细胞种群约为105 106的当前商业电穿孔设备中，这很难实现。我们计划设计，构建和表征批处理型纳米型纳米式夹层夹层电穿孔设备（NSE）设备和连续型收敛流动电动型（CFE）设备。然后，这两个新型设备将用于研究体外转染效率，并吸收选定的寡核苷酸和用于癌症和干细胞应用的基因。它们的性能将与最佳的商业电穿孔系统进行比较。将开发几种新型的电穿孔设备，以更好地体外和体外转染治疗材料。在这项研究中，将研究寡核苷酸和质粒DNA对癌细胞和干细胞的摄取。所提出的设备具有将寡核苷酸和质粒DNA转染的巨大潜力，以用于临床应用。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Lipid nanoparticles for hepatic delivery of small interfering RNA.

DOI：
10.1016/j.biomaterials.2012.05.002
发表时间：
2012-09
期刊：
BIOMATERIALS
影响因子：
14
作者：
Yu, Bo;Hsu, Shu-Hao;Zhou, Chenguang;Wang, Xinmei;Terp, Megan C.;Wu, Yun;Teng, Lesheng;Mao, Yicheng;Wang, Feng;Xue, Weiming;Jacob, Samson T.;Ghoshal, Kalpana;Lee, Robert J.;Lee, Ly J.
通讯作者：
Lee, Ly J.

Semicontinuous flow electroporation chip for high-throughput transfection on mammalian cells.