In vivo monitoring of cellular signaling and metabolite flux is an essential tool to study the dynamics of cellular networks and the effects of re-engineering these networks.
The aims of this proposal are the development and use of a modular, tunable, allosterically controlled biosensor platform that can simultaneously monitor and re-engineer cellular pathways. Small, noncoding RNA molecules (allosteric ribozymes, riboswitches and antiswitches) will be the basis of these biosensors and will be developed to control a fluorescent resonant energy transfer event in response to small molecule ligands or cellular signalling events. In addition, sensor components will be engineered to allow for simultaneous control of expression of endogenous cellular transcripts. A mitogen activated protein kinase pathway from S. cerevisiae will serve as a model system for biosensor development, adaptation, and translational control. These studies are predicted to establish a biosensor platform with rapid detection kinetics that is easily evolvable, modular, and capable of allosteric control of mRNA expression. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM078901-03
Application #
7457761
Study Section
Special Emphasis Panel (ZRG1-F14-A (20))
Program Officer
Fabian, Miles
Project Start
2006-07-01
Project End
2009-06-30
Budget Start
2008-07-01
Budget End
2009-06-30
Support Year
3
Fiscal Year
2008
Total Cost
$53,992
Indirect Cost
Name
California Institute of Technology
Department
Type
Schools of Engineering
DUNS #
009584210
City
Pasadena
State
CA
Country
United States
Zip Code
91125
Hoff, Kevin G; Culler, Stephanie J; Nguyen, Peter Q et al. (2009) In vivo fluorescent detection of Fe-S clusters coordinated by human GRX2. Chem Biol 16:1299-308
Hoff, Kevin G; Goodlitt, Rochelle; Li, Rui et al. (2009) Fluorescence detection of a protein-bound 2Fe2S cluster. Chembiochem 10:667-70