Multiple functionalities of functional nucleic acids for developing high-performance lateral flow assays

Functional nucleic acids (FNAs) have emerged as superior molecular recognition elements with high binding affinity, specificity, and catalytic activity. Due to multiple functionalities like specific recognition, stimulus responsiveness, sequence programmability, structural tailorability, and ease of modification, FNAs have been integrated into lateral flow assays (LFAs) to construct simple and sustainable paper-based chromatographic biosensors for rapid, antibody-free, cost-effective, and user-friendly target detection. This review presented a systematic, comprehensive summary of functional nucleic acid-based lateral flow assays (FNA-LFAs). First, the screening, characterization, and tailoring of FNAs to meet the requirements of LFAs were briefly introduced. Second, the multi-functionality of FNAs for developing high-performance LFAs is presented, along with the entire LFA process from sample pretreatment to signal transduction. Finally, the applications of FNA-LFAs in point-of-care testing (POCT) are illustrated along with future research perspectives, aiming to provide a valuable reference for researchers in the field of FNA sensing. • Provided a systematic and comprehensive summary of FNA-LFAs. • Listed and further analyzed six types of FNAs applied in LFAs. • Highlighted the multiple functionalities of FNAs for developing high-performance LFAs. • Dissected the entire LFA construction process step-by-step. • Introduced the primary FNA-LFA applications areas in POCT in detail.