IEC 195 |
| Gang Bao1, Andrew Tsourkas2, and Phil Santangelo1. (1) Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 315 Ferst Dr, Suite 2306, Atlanta, GA 30332, (2) MGH-CMIR, Massachusetts General Hospital and Harvard Medical School, Building 149, 13th Street, Room 5404, Charlestown, MA 02129-2060 |
| Molecular beacons provide a simple and promising tool for the detection of target mRNA due to their ability to differentiate between bound and unbound probes with high signal-to-noise ratio and improved specificity. However, the harsh intracellular environment does limit the detection sensitivity in vivo. To overcome this difficulty, we have developed a dual FRET molecular beacons approach that utilizes a pair of molecular beacons with FRET dyes. When both molecular beacons are bound to adjacent sites on the same target mRNA, there is an increase in acceptor fluorescence upon donor excitation due to FRET. Since FRET is extremely sensitive to distance between donor and acceptor molecules, it occurs only when the donor and acceptor moleculars are both bound to the same mRNA target. Therefore, detecting fluorescence due to FRET can significantly reduce signal contamination from beacon degradation and spontaneous opening. To better understand and optimize the behavior of molecular beacons we have systematically studied the hybridization kinetics and thermodynamics of molecular beacons with structural variations. It was found that by adjusting the stem and probe length the kinetics and specificity of molecular beacons could be tailored for a particular application. Various methods were also employed to improve the signal-to-noise ratio of the dual FRET molecular beacons. |
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Biological Applications of Nanomaterials and Nanotechnology (sponsored by Advanced Materials & Nanotechnology Subdivision)
8:00 AM-12:05 PM, Wednesday, March 26, 2003 Convention Center -- Room 393, Oral
Division of Industrial and Engineering Chemistry |