Thesis

Filling the GAPS in Neurofibromatosis type 1

Neurofibromatosis type 1 (NF1) is an autosomal dominant disorder that greatly impacts society because its high frequency. Its symptoms are diverse and occur without prior warning. Symptoms include neural tumors, which may result in loss of vision or other senses, and symptoms differ between individuals with identical NF1 mutations. This tumor suppressor gene produces a protein called Neurofibromin, which has arginine fingers and is able to aid in the hydrolysis of RAS-GTP to RAS-GDP. The GAP domain of Neurofibromin was cloned into a mammalian expression vector at position 1300, outside the multiple cloning site in this vector. Upon transfection of a NF1-/- fibroblast, a colorimetric assay was performed to investigate how much RAS-GTP was present in cells transfected with the construct. A significant difference in RAS-GTP levels (p<0.05) suggests that the GAP domain of Neurofibromin was successfully transcribed on active RAS. These results may open the way for creating a "Trojan Horse", which could cross the blood brain barrier, allowing for treatment of disorders that affect the central nervous system. In NF1, this treatment could prevent Optic Pathway Gliomas, a complication of NF1. In preparation for the creation of a "Trojan Horse", the multiple cloning site in this construct could be used to investigate lactotransferrin, which has potential for receptor-mediated transcytosis

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