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.
| Reinhart, Peter H; Kaltenbach, Linda S; Essrich, Christian et al. (2011) Identification of anti-inflammatory targets for Huntington's disease using a brain slice-based screening assay. Neurobiol Dis 43:248-56 |
| Hoffstrom, Benjamin G; Kaplan, Anna; Letso, Reka et al. (2010) Inhibitors of protein disulfide isomerase suppress apoptosis induced by misfolded proteins. Nat Chem Biol 6:900-6 |
| Braithwaite, Steven P; Schmid, Ralf S; He, Dong Ning et al. (2010) Inhibition of c-Jun kinase provides neuroprotection in a model of Alzheimer's disease. Neurobiol Dis 39:311-7 |
