Journal
ACS APPLIED BIO MATERIALS
Volume 3, Issue 5, Pages 2633-2642Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsabm.9b01237
Keywords
aptamer; biosensors; cellular imaging; fluorogenic RNA; ratiometric sensors
Funding
- UMass Amherst startup grant
- NIH [RO1A1136789]
- NSF
- Alfred P. Sloan Research Fellowship
Ask authors/readers for more resources
Imaging the cellular dynamics of metabolites and signaling molecules is critical for understanding various metabolism and signal transduction pathways. Genetically encoded RNA-based sensors are emerging powerful tools for this purpose. However, it was challenging to use these sensors to precisely determine the intracellular concentrations of target analytes. To solve this problem, we have recently developed ratiometric sensors using an orthogonal pair of RNA/fluorophore conjugates: Broccoli/DFHBI-1T (3,5-difluoro-4-hydroxybenzylidene-1-trifluoroethyl-imidazolinone) and DNB (dinitroaniline-binding aptamer)/SR-DN (sulforhodamine B-dinitroaniline). The cellular DNB-to-Broccoli fluorescence intensity ratio can be directly applied to quantify the target concentrations at the single-cell level. Unfortunately, due to the instability of the SR-DN dye, this ratiometric sensor is difficult to use for monitoring target dynamics. Herein, by replacing SR-DN with a stable TMR (tetramethylrhodamine)-DN dye, we developed a ratiometric sensor system based on Broccoli/DFHBI-1T and DNB/TMR-DN, which can be used for dynamic imaging in living cells. We believe these advanced genetically encoded ratiometric sensors can be widely used for intracellular studies of various target analytes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available