A dual-signal biosensor based on surface-enhanced Raman spectroscopy for high-sensitivity quantitative detection and imaging of circRNA in living cells

Circular RNAs (circRNAs) are non-coding RNAs that play key roles in the development and progression of cancer through various mechanisms of action, making them promising biomarkers for cancer diagnosis, prognosis, and treatment. In the present study, a biosensor based on surface-enhanced Raman spectroscopy (SERS) was developed for rapid, simple, and sensitive quantitative detection of intracellular circRNAs for the first time. A dual-signal SERS nanoprobe with a 4MBN and ROX signal molecule was fabricated, and the ROX signal intensity was used to determine the concentration of target circSATB2. 4MBN was used as an internal standard to calibrate the ROX signal, thereby achieving highly sensitive and reliable detection of the target circRNA with a limit of detection of 0.043 pM. Furthermore, the relatively high expression of circSATB2 in lung cancer cells compared to that in normal lung epithelial cells was successfully characterized by the proposed SERS imaging method, which is consistent with the results of standard reverse transcription-polymerase chain reaction (RT-PCR). Monitoring of specific circRNAs using this SERS-based biosensor is a promising method for cancer diagnosis and gene therapy.