BMEBT DISSERTATION PROPOSAL PRESENTATION
When: Wednesday,
January 31, 2018
10:00 AM
-
11:00 AM
Where: > See description for location
Description: The Biomedical Engineering and Biotechnology (BMEBT) Program is pleased to announce a dissertation proposal presentation by AJ Vincelli. Snacks will be provided.
Proposal Title: Engineering a Platform Technology to Incorporate a Fluorescent Amino Acid into Proteins for Live-Cell Imaging of Mistrafficking Events
Abstract: Thousands of diseases are thought to be caused by genetic mutations and subsequent aberrant protein localization within the cell, though pathological mechanisms are understood for only a fraction of all known mutations. This project aims to engineer a widely-applicable platform which can visualize real-time protein trafficking events in living human cells. First, a mutant library of yeast tryptophanyl-tRNA synthetases (RS) will be rationally designed to recognize a small coumarin fluorescent amino acid. The yeast enzyme is thought to be orthogonal to human tryptophan aminoacylation processes, yet still recognizable to the mammalian cell's translational machinery. Second, a cognate yeast amber suppressor coumarinyl tRNA will also be engineered, which will be charged with free coumarin amino acid by the RS. When cloned into a vector and transfected into human cells, these engineered biomolecules will insert the small coumarin fluorescent amino acid into a desired position within a protein of interest without perturbing its native structure or function. After confirming platform functionality in human cells using a GFP reporter assay, this technology will be applied to a proof-of-concept mistrafficking disease model. A single missense mutation in the protein of interest abolishes its localization signal, and thus it accumulates in the incorrect cellular compartment and causes disease. Both the wild-type protein trafficking, as well as mutant protein localization, will be visualized in real time using confocal microscopy. This proposed novel platform technology has broad applicability to protein localization studies which can reveal the pathogenesis of mistrafficking pathways, validate suspected mistrafficking mutations, and enable a greater understanding of disease-related phenotypes at the subcellular level.
Location: SENG-311
RSVP To: AJ Vincelli, avincelli@umassd.edu (please RSVP, so we know how much food to bring!)
Advisor: Dr. Maolin Guo
Committee Members: Dr. Tracie Ferreira, Dr. Xiaofei Jia, and Dr. Firas Khatib
Proposal Title: Engineering a Platform Technology to Incorporate a Fluorescent Amino Acid into Proteins for Live-Cell Imaging of Mistrafficking Events
Abstract: Thousands of diseases are thought to be caused by genetic mutations and subsequent aberrant protein localization within the cell, though pathological mechanisms are understood for only a fraction of all known mutations. This project aims to engineer a widely-applicable platform which can visualize real-time protein trafficking events in living human cells. First, a mutant library of yeast tryptophanyl-tRNA synthetases (RS) will be rationally designed to recognize a small coumarin fluorescent amino acid. The yeast enzyme is thought to be orthogonal to human tryptophan aminoacylation processes, yet still recognizable to the mammalian cell's translational machinery. Second, a cognate yeast amber suppressor coumarinyl tRNA will also be engineered, which will be charged with free coumarin amino acid by the RS. When cloned into a vector and transfected into human cells, these engineered biomolecules will insert the small coumarin fluorescent amino acid into a desired position within a protein of interest without perturbing its native structure or function. After confirming platform functionality in human cells using a GFP reporter assay, this technology will be applied to a proof-of-concept mistrafficking disease model. A single missense mutation in the protein of interest abolishes its localization signal, and thus it accumulates in the incorrect cellular compartment and causes disease. Both the wild-type protein trafficking, as well as mutant protein localization, will be visualized in real time using confocal microscopy. This proposed novel platform technology has broad applicability to protein localization studies which can reveal the pathogenesis of mistrafficking pathways, validate suspected mistrafficking mutations, and enable a greater understanding of disease-related phenotypes at the subcellular level.
Location: SENG-311
RSVP To: AJ Vincelli, avincelli@umassd.edu (please RSVP, so we know how much food to bring!)
Advisor: Dr. Maolin Guo
Committee Members: Dr. Tracie Ferreira, Dr. Xiaofei Jia, and Dr. Firas Khatib
Contact: > See Description for contact information
Topical Areas: Students, Graduate, Chemistry and Biochemistry, Bioengineering, Computer and Information Science, Research, Sponsored Projects Administration