Exploring a Novel Technique to Engineer Donor Cells for Transplantation Following Neurotoxin Exposure.

摘要

Loss of neurons due to neurotoxin exposure result in various forms of neurodegenerative disorders (ND). Promising strategies include transplantation of donor cells The quantity and quality of donor cells however has been a problem. We have been exploring DNA molecular decoy as a way to engineer donor cells that are suitable for transplantation. We have been using a double stranded (ds) DNA molecule that includes the septamer motif (Dobi et al., 2000). According to Objective (star)1: we have determined that (a) the optimal developmental stage for decoy using the rat brain is between El6 and E20; (b) the duration of 4 days of decoy is sufficient for generating BrdU+/nestin+ cells; (c) the decoy molecule is sufficiently stabile without additional modifications, (d) using DNA delivery systems (PEI or DOTAP) for increased delivery of dsDNA decoy molecules increased toxicity. In addition, we have determined that the septamer decoy molecule specifically interact with the septamer nuclear complex without interfering with other DNA binding regulatory proteins; established a fast and easy assay system for testing decoy molecules; determined that septamer DNA decoy decreases the number of postmitotic (MAP2+) neurons, suggesting a potential 'dedifferentiation'. The differentiation potential of decoyed cells (Objective No. 2) are to be tested in the next set of experiments.