QD-FRET-based biosensing of small molecule analytes using transcription factor-DNA binding

Thuy T. Nguyen; Margaret Chern; R. C. Baer; Andy Fan; Chloe Grazon; James Galagan; Allison M. Dennis

doi:10.1117/12.2516576

22 May 2019 QD-FRET-based biosensing of small molecule analytes using transcription factor-DNA binding

Thuy T. Nguyen, Margaret Chern, R. C. Baer, Andy Fan, Chloe Grazon, James Galagan, Allison M. Dennis

Proceedings Volume 10892, Colloidal Nanoparticles for Biomedical Applications XIV; 108920Q (2019) https://doi.org/10.1117/12.2516576
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

An alternative molecular recognition approach was developed for sensing small molecule analytes using the differential binding of an allosteric transcription factor (TF, specifically TetR) to its cognate DNA as the molecular recognition element coupled with fluorescent resonance energy transfer (FRET) to yield an internally calibrated optical signal transduction mechanism. Sensors were evaluated comprising Cy5-modified DNA (FRET acceptor) with either a tdTomato-TetR fusion protein (FP-TF) or quantum dot-TetR conjugate (QD-TF) as the FRET donor by measuring the ratio of acceptor and donor fluorescence intensities (FA/FD) with titrations of a derivative of the antibiotic tetracycline, anhydrous tetracycline (aTc). A proof-of-concept FRET-based biosensor was successfully demonstrated through the modulation of FA/FD signal intensities based on varying analyte concentrations. Sensor design parameters affecting overall signal-to-noise ratio and sensitivity of the sensors are also identified.

Citation Download Citation

Thuy T. Nguyen, Margaret Chern, R. C. Baer, Andy Fan, Chloe Grazon, James Galagan, and Allison M. Dennis "QD-FRET-based biosensing of small molecule analytes using transcription factor-DNA binding", Proc. SPIE 10892, Colloidal Nanoparticles for Biomedical Applications XIV, 108920Q (22 May 2019); https://doi.org/10.1117/12.2516576

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available