Functional Analysis of Neurofibromin: Clues from Drosophila Applied to Mammalian Systems
Abstract
The purpose of our research is the generation of relevant biological assay systems in which Ras-independent effects of neurofibromin on cellular proliferation can be readily assessed. Our primary efforts have been aimed at overcoming the longstanding difficulties of manipulating normal and mutant forms of neurofibromin in mammalian cells. We have focused on the development of tightly controlled expression systems using both a vector-based ecdysone system and a tetracycline-regulated HSV amplicon system in established NIH3T3 murine fibroblasts and primary neurofibromin-deficient mouse embryo fibroblasts. Our progress has been severely limited by technical difficulties with both systems that have prevented the establishment of reliably inducible exogenous neurofibromin expression in either cell type. We are now utilizing a recently described retroviral based ecdysone expression system since stable trans-activator-expressing NIH3T3 lines in this system are available. Additionally, expanded transgene delivery into poorly transfected cell lines is afforded by retrovirus infection. These important features will overcome the two major obstacles that have hampered our progress to date. The revised experiments outlined in this report will lay the foundation for further investigation of the complex role of neurofibromin in cellular growth control mediated through its Ras-dependent and Ras-independent functions.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 2001
- Accession Number
- ADA400501
Entities
People
- Rosemary Foster
Organizations
- Massachusetts General Hospital