Gold-Nanobeacons as a theranostic system for the detection and inhibition of specific genes

Abstract

This protocol describes the synthesis and detailed calibration of a gold nanoparticle-based nanobeacon \ (Au-nanobeacon) as an innovative theranostic approach for detection and inhibition of sequence-speci c DNA and RNA for in vitro and ex vivo applications. Under hairpin con guration, proximity to gold nanoparticles leads to uorescence quenching; hybridization to a complementary target restores uorescence emission due to the gold nanobeacons' conformational reorganization that causes the uorophore and the AuNP to part from each other. This concept can easily be extended and adapted to assist the in vitro evaluation of silencing potential of a given sequence to be later used for ex vivo gene silencing and RNAi approaches, with the ability to monitor real-time gene delivery action. The time range for the entire protocol is ~8 days, including synthesis, functionalization and calibration of Aunanobeacons, RNAi and gene silencing assays.