High-Efficiency Biolistic Device for Brain Transfection

用于脑转染的高效基因枪装置

• 批准号: 7010713
• 负责人: DONALD C LO
• 金额: $ 17.39万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7010713
• 关键词: DNA; albino rat; biomedical equipment development; biotechnology; brain; gene delivery system; gene expression; high throughput technology; immature animal; sectioning; tissue /cell culture; transfection

device, based on particle-mediated gene transfer (or 'biolistics'), for the identification and validation of candidate therapeutic drugs and targets. The tremendous pace at which new genomic and proteomic technologies are advancing and generating new information regarding the normal and pathological brain has created a major bottleneck in translational neuroscience, namely, the ability to physiologically validate new candidate therapeutic targets upon which to base target-driven efforts in medicinal chemistry. In this context, a major technological rate-limiting step has been the difficulty of manipulating the expression of genes and proteins in neural disease-specific experimental systems based on bona fide, postmitotic neurons, ranging from primary cultures to tissue explant models. In preliminary experiments, we have demonstrated that a neural transfection approach based on particle-mediated gene transfer, or biolistics, can be used to screen DMA-based and chemical libraries at medium-throughput levels in disease-specific models created in living brain slices. We propose here to develop a new particle acceleration-based technology to increase transfection efficiency and throughput by 10-fold or more. Such an improved and affordable biolistic device would allow laboratory groups of standard size to embark upon high-content, functional screening and validation experiments in neural cells and tissues using the mass of genomic and proteomic information now available. The Research Plan has 3 specific aims: Specific Aim1: To develop a new and highly efficient particle-acceleration transfection device. Specific Aim 2: To determine workable ranges of shooting parameters for the new device. Specific Aim 3: To evaluate the efficiency and effectiveness of the new biolistic device with respect to existing technology.

基于粒子介导的基因转移(或“生物生物学”)的设备，用于识别和验证候选治疗药物和靶点。新的基因组和蛋白质组技术的进步和产生关于正常和病理大脑的新信息的巨大速度已经在翻译神经科学中产生了一个主要的瓶颈，即从生理上验证新的候选治疗靶点的能力，以此为基础的药物化学的靶向驱动的努力。在这种背景下，一个主要的技术限制步骤是在基于真正的有丝分裂后神经元的神经疾病特异性实验系统中操纵基因和蛋白质的表达的困难，从原代培养到组织外植体模型。 在初步的实验中，我们已经证明了一种基于 颗粒介导的基因转移，或称生物生物学，可用于在活体脑片创建的疾病特异性模型中以中等吞吐量水平筛选基于DMA的库和化学库。我们建议开发一种新的基于粒子加速的技术，将转染率和吞吐量提高10倍或更多。这样一种改进的、负担得起的生物化学设备将允许标准规模的实验室小组利用现有的大量基因组和蛋白质组信息，在神经细胞和组织中开展高含量、功能筛选和验证实验。《研究计划》有3个具体目标： 目的：研制一种新型高效的粒子加速转染器。 具体目标2：确定新设备可操作的射击参数范围。 具体目标3：评估新的生物装置相对于现有技术的效率和效果。

项目成果

Identification of anti-inflammatory targets for Huntington's disease using a brain slice-based screening assay.

使用基于脑切片的筛选测定来鉴定亨廷顿病的抗炎靶标。

• DOI: 10.1016/j.nbd.2011.03.017
• 发表时间: 2011
• 期刊: Neurobiology of disease
• 影响因子: 6.1
• 作者: Reinhart,PeterH;Kaltenbach,LindaS;Essrich,Christian;Dunn,DeniseE;Eudailey,JoshuaA;DeMarco,CTodd;Turmel,GregoryJ;Whaley,JenniferC;Wood,Andrew;Cho,Seongeun;Lo,DonaldC
• 通讯作者: Lo,DonaldC

Inhibition of c-Jun kinase provides neuroprotection in a model of Alzheimer's disease.

• DOI: 10.1016/j.nbd.2010.04.015
• 发表时间: 2010-09
• 期刊: Neurobiology of disease
• 影响因子: 6.1
• 作者: Braithwaite SP;Schmid RS;He DN;Sung ML;Cho S;Resnick L;Monaghan MM;Hirst WD;Essrich C;Reinhart PH;Lo DC
• 通讯作者: Lo DC