Dissertation/Thesis Abstract

Bioengineering Novel Reporter Proteins
by Bartelle, Benjamin B., Ph.D., New York University, 2013, 225; 3556976
Abstract (Summary)

Visualization of gene expression has led to a revolution in biology over the past two decades. Primarily this visualization has occurred using fluorescent proteins, like GFP, that can be directly visualized with microscopy. Fluorescence imaging is limited by depth of penetration when applied to living mice or humans however. For this, MRI, ultrasound and other modalities are under continual development for in vivo applications. Ideally, every in vivo imaging modality would have their own reporter genes, allowing for unconstrained genetic studies of structure and function. The current wealth of bioinformatics data presents a rich pallet of starting materials for bioengineering this next generation of reporter proteins.

This work utilized multiple approaches to creating reporters: cell labeling with, "Biotag" derived from a bacterial biotinylation enzyme and substrate; genetically controlled absorption of the MRI contrast agent Mn via the metal transport protein DMT1; and sequestration of Mn using the metal sensing transcription factor MntR. The reporter proteins were implemented in tissue culture and living mice to give a new view of gene expression in processes such as neural and vascular development. Moreover, the development process yielded new insights into the proteins themselves and the context in which they function. Each method has particular strengths and limitations but are, at present, the vanguard of in vivo molecular imaging.

Indexing (document details)
Advisor: Turnbull, Daniel H.
Commitee: Canary, James W., Johnson, Glyn, Joyner, Alexandra L., Stokes, David L.
School: New York University
Department: Basic Medical Science
School Location: United States -- New York
Source: DAI-B 74/07(E), Dissertation Abstracts International
Source Type: DISSERTATION
Subjects: Molecular biology, Biomedical engineering, Biophysics
Keywords: Biomedical imaging, In vivo imaging, Mri, Protein engineering, Reporter genes, Reporter proteins, Ultrasound
Publication Number: 3556976
ISBN: 9781267995520
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