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Bioengineering Novel Reporter Proteins

<p> 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 <i>in vivo</i> applications. Ideally, every <i> in vivo</i> 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. </p><p> 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 <i>in vivo</i> molecular imaging. </p>

Identiferoai:union.ndltd.org:PROQUEST/oai:pqdtoai.proquest.com:3556976
Date25 April 2013
CreatorsBartelle, Benjamin B.
PublisherNew York University
Source SetsProQuest.com
LanguageEnglish
Detected LanguageEnglish
Typethesis

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