Nervous and nonnervous cell transduction by recombinant adenoviruses that inducibly express the human PrP

The study of the prion protein (PrP) physiological functions or its specifi c role in transmissible spongiform encephalopathies (TSE) requires new tool s, particularly those able to induce PrP overexpression in a large range of cells, in vivo as well as in vitro. Here we describe the construction of t wo recombinant adenoviruses encoding the human PrP either with a valine at position 129 (AdTRVal) or a methionine (AdTRMet). Both genes were put under the control of the tetracycline-responsive promoter, allowing tight regula tion of PrP expression. AdTRVal and AdTRMet induced high expression of the human PrP in CHO-KI cells and in organotypic brain slices in culture. The p roteins expressed from these viruses exhibited a glycosylphosphatidyl inosi tol (GPI) anchor, proper glycosylation and sensitivity to proteinase K dige stion. AdTRVal and AdTRMet will allow future studies on the human PrP and o n the role of the codon 129 polyphormism in human TSE. (C) 2001 Academic Pr ess.