In this administrative supplement, we request funds for a new instrument, Molecular Devices ImageXpress Micro 4 High Content Screening System. The instrument will allow for the measurement of both luminescence and fluorescence of tailored fluorescent and chemiluminescent Fe(II) and H2O2 responsive chemical inducers in chemical systems and in live cell and subcellular localization imaging and quantitative detection of luminescence and fluorescence. In the funded 1R01GM130772 grant, we propose a novel synthetic biology technology termed ?Environment Stimuli-Induced Proximity (ESIP)? to spatiotemporally manipulate cellular functions. To demonstrate the feasibility, we propose to develop fluorescent and chemiluminescent Fe(II) and H2O2 responsive ESIP chemical inducers. Specifically, we will design, synthesize, test and optimize Fe2+ and H2O2-responsive fluorescent and chemiluminescent ESIP inducers (Specific Aims 1-2) and test them in cells and construct ESIP-mediated ?AND? Boolean logic gates to control the conditional production of AD therapeutic proteins (Specific Aim 3). Therefore, this instrument is crucial for us to quickly and accurately characterize and detect fluorescent and chemiluminescent Fe(II) and H2O2 responsive ESIP chemical inducers in chemical and biological systems. This instrument will be the first at the University of Arizona, and will be available to all the Principal Investigators who are currently funded by NIGMS and other mechanisms.
We wish to develop a new synthetic biology strategy, termed ?environmental stimuli-induced proximity (ESIP)? by developing `smart' fluorescent and chemiluminescent chemical inducers to spatiotemporally control cellular signaling and functions. The bifunctional probes will selectively sense cellular signal molecules (e.g., Fe2+ and H2O2) to produce original bioactive inducers with spatiotemporal precision to induce downstream biological events. The technology will ultimately be transformed into entirely new cell/gene therapies for disease treatment such as neurodegenerative diseases with fewer side effects.
