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:
- 金额:$ 20.62万
- 依托单位:
- 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:
- 关键词:AcademiaAccelerationAcidsAreaBiological ModelsCell CountCell LineCell SurvivalCell membraneCellsClinicalClinical TrialsConditionCultured CellsDNADevicesDigit structureElectroporationEncapsulatedEvaluationGene DeliveryGene TransferGenesGovernmentGrowth FactorIn VitroLocalizedMarketingMelanoma CellMembraneMethodsMicroRNAsMicrocapsules drug delivery systemMicrofluidicsMolecular ProbesMotionMusNuclearNucleic AcidsOligonucleotidesOperative Surgical ProceduresPatientsPerformancePersonsPharmacologic SubstancePharyngeal structurePhysiologic pulsePolymersPopulationProcessProviderPulse takingPurposeRangeReporter GenesResearchResearch ProposalsSmall Interfering RNASolutionsStem cellsSystemT-LymphocyteTechniquesTechnologyTestingTherapeuticTimeTissuesTransfectionViralWorkbaseblastomere structurecancer cellcancer stem cellclinical applicationdesign and constructiondesireelectric fieldembryonic stem cellexperiencein vivoknock-downleukemiananofluidicnovelplasmid DNAtooltranscription factortwo-dimensionaluptakevoltage
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。最近的进展使得该方法适用于广泛的细胞和生物分子,用于体外转染测试和离体甚至体内临床试验。目前全球电穿孔市场约为2500万美元,预计在学术界、政府、制药和生物技术公司的刺激下,未来五年将以两位数的增长迅速增长。目前商业化的电穿孔装置和试剂盒(特别是Amaxa Nucleofector技术)是方便的、易于使用的,并且当需要非病毒基因转移时对于许多难以转染的细胞系是有益的。然而,一个主要的缺点是,每个新的细胞系统(如实验室特异性克隆或患者的原代细胞)通常需要耗时且昂贵的试错搜索过程来鉴定适当的电穿孔条件和/或溶液组成,其将实现具有足够高的细胞活力的期望转染水平。每个电池系统必须使用精细调谐的高电压和非均匀电场单独优化。如果电穿孔可以在温和、均匀的电场中进行,并且对各种细胞系和原代细胞都有效,这将是非常有价值的。通过这种方式,可以消除或高度简化为每个单元系统确定新设置的需要。对于临床应用,需要以高转染效率和细胞活力转染大量细胞(例如>109个细胞)。这在推荐的细胞群体为约105的当前商业电穿孔装置中难以实现 106.我们计划设计,构建和表征一个批量型纳米喷嘴三明治电穿孔(NSE)设备和一个连续型会聚流电穿孔(CFE)设备。这两种新型装置将用于研究体外转染效率和癌症和干细胞应用中选定的寡核苷酸和基因的摄取。它们的性能将与最好的商业电穿孔系统进行比较。为了更好地在体外和离体转染治疗材料,将开发几种新型电穿孔装置。在这项研究中,将研究寡核苷酸和质粒DNA进入癌细胞和干细胞的摄取。所提出的装置具有将寡核苷酸和质粒DNA转染到大量细胞中用于临床应用的巨大潜力。
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:
- 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.
Targeted nanoparticles enhanced flow electroporation of antisense oligonucleotides in leukemia cells.
- DOI:10.1016/j.bios.2010.06.025
- 发表时间:2010-10-15
- 期刊:
- 影响因子:12.6
- 作者:Wang S;Zhang X;Yu B;Lee RJ;Lee LJ
- 通讯作者:Lee LJ
Semicontinuous flow electroporation chip for high-throughput transfection on mammalian cells.
- DOI:10.1021/ac9002672
- 发表时间:2009-06-01
- 期刊:
- 影响因子:7.4
- 作者:Wang, Shengnian;Zhang, Xulang;Wang, Weixiong;Lee, L. James
- 通讯作者:Lee, L. James
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Ly James Lee其他文献
Ly James Lee的其他文献
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{{ truncateString('Ly James Lee', 18)}}的其他基金
Multi-parametric Integrated Molecular Detection of SARS-CoV-2 from Biofluids by Adapting Single Extracellular Vesicle Characterization Technologies
采用单细胞外囊泡表征技术对生物体液中的 SARS-CoV-2 进行多参数集成分子检测
- 批准号:
10266279 - 财政年份:2020
- 资助金额:
$ 20.62万 - 项目类别:
Extracellular Vesicles in Small Cell Lung Cancer Early Detection
小细胞肺癌早期检测中的细胞外囊泡
- 批准号:
10115627 - 财政年份:2017
- 资助金额:
$ 20.62万 - 项目类别:
Large Scale Nanochannel Electroporation (NEP) for Cell Reprogramming
用于细胞重编程的大规模纳米通道电穿孔 (NEP)
- 批准号:
8583897 - 财政年份:2013
- 资助金额:
$ 20.62万 - 项目类别:
Plasma RNA based Early Lung Cancer Detection by Tethered Cationic Lipoplex Assay
通过系留阳离子脂质复合物检测进行基于血浆 RNA 的早期肺癌检测
- 批准号:
8570641 - 财政年份:2013
- 资助金额:
$ 20.62万 - 项目类别:
Plasma RNA based Early Lung Cancer Detection by Tethered Cationic Lipoplex Assay
通过系留阳离子脂质复合物检测进行基于血浆 RNA 的早期肺癌检测
- 批准号:
8735903 - 财政年份:2013
- 资助金额:
$ 20.62万 - 项目类别:
Large Scale Nanochannel Electroporation (NEP) for Cell Reprogramming
用于细胞重编程的大规模纳米通道电穿孔 (NEP)
- 批准号:
8702172 - 财政年份:2013
- 资助金额:
$ 20.62万 - 项目类别:
Large Scale Nanochannel Electroporation (NEP) for Cell Reprogramming
用于细胞重编程的大规模纳米通道电穿孔 (NEP)
- 批准号:
8774717 - 财政年份:2013
- 资助金额:
$ 20.62万 - 项目类别:
A Renewal Proposal for the Nanoscale Science and Engineering Center (NSEC) for Affordable Nanoengineering of Polymeric Biomedical Devices
纳米科学与工程中心 (NSEC) 的更新提案,以实现经济实惠的聚合物生物医学设备纳米工程
- 批准号:
0914790 - 财政年份:2009
- 资助金额:
$ 20.62万 - 项目类别:
Cooperative Agreement
Novel Micro/nanofluidic Electroporation Devices for DNA&Oligonucleotide Delivery
新型 DNA 微/纳流体电穿孔装置
- 批准号:
7363207 - 财政年份:2007
- 资助金额:
$ 20.62万 - 项目类别:
NSEC: Center for Affordable Nanoengineering of Polymer Biomedical Devices (CANPBD)
NSEC:经济实惠的聚合物生物医学设备纳米工程中心 (CANPBD)
- 批准号:
0425626 - 财政年份:2004
- 资助金额:
$ 20.62万 - 项目类别:
Cooperative Agreement
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