The research goal of the Vector Development Section is to design vectors which overcome problems inherent to current gene transfer approaches such as transient expression (e.g. lipid-mediated plasmid DNA transfer, adenovirus vectors) and the risk for random integration and potential mutagenesis (e.g. retrovirus, adeno-associated virus). An optimal approach would be a vector that provides: (1) stable gene transfer without integration into the host genome, and (2) unlimited capacity to carry DNA so that full length genes can be transferred and analyzed functionally. Toward this end, we are attempting to construct self replicating-vectors in the form of human artificial chromosomes (HAC) which contain human DNA sequences likely necessary for proper chromosome function including telomeres, centromeres and replication origins (Project 1). In addition to the construction of these large vectors, we are also attempting to optimize the delivery of high molecular weight DNA to mammalian cells (Project 2). The primary focus for this study is the cystic fibrosis transmembrane conductance regulator (CFTR) gene contained within in a yeast artificial chromosome (YAC) which also expresses a marker gene Ecoli Lac Z gene [coding for beta-galactosidase]. We are now applying the strategy developed for the CFTR YAC to other YACs and other genetic diseases (Project 3).
