High-Efficiency Biolistic Device for Brain Transfection

基于颗粒介导的基因转移（或“生物学”）的设备，用于鉴定和验证候选治疗药物和靶标。新的基因组学和蛋白质组学技术正在飞速发展，并产生了关于正常和病理大脑的新信息，这给转化神经科学带来了一个主要的瓶颈，即在生理上验证新的候选治疗靶点的能力，从而在药物化学中建立靶点驱动的努力。在这种情况下，一个主要的技术限制步骤是在基于真实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表达的困难，从原代培养到组织外植体模型。

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.