Functional Analysis of Neurofibromin: Clues From Drosophila Applied to Mammalian Systems
Abstract
The purpose of our research was the generation of relevant biological assay systems in which Ras-independent effects of neurofibromin on cellular proliferation could be assessed. Our primary efforts have been aimed at overcoming longstanding difficulties of manipulating normal and mutant forms of neurofibromin in mammalian cells. We initially focused on development of tightly controlled expression systems using vector- and retroviral-based ecdysone systems and a tetracycline-regulated amplicon system in established NIH3T3 murine fibroblasts and neurofibromin-deficient primary mouse embryo fibroblasts (MEFs). Our progress has been limited by technical difficulties with all three systems that prevented establishment of reliably inducible exogenous neurofibromin expression in either cell type. We have utilized a recently developed modified tetracycline-regulated amplicon system that affords tighter control of exogenous gene expression. Our preliminary analyses in NIH3T3 fibroblasts, neurofibromin-deficient MEFs, and human Schwann cells derived from NFl-associated tumors show improved control of tetracycline-inducible gene expression and suggest that this amplicon system will be a valuable tool in analysis of neurofibromin function in a broad spectrum of cell types, including those that are pathologically relevant to the NFl phenotype. We expect that further studies utilizing the amplicon system will help to clarify the complex role of neurofibromin in cellular growth control.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 2002
- Accession Number
- ADA412174
Entities
People
- Rosemary Foster
Organizations
- Massachusetts General Hospital