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From the first investigations on peptide nucleic acids (PNA), peculiar features have been described as intrinsic of the singular chemical nature of these biomolecules. Relevant applications in the development of biosensors occurred in the last decades, exploiting them as bioreceptors. Their outstanding chemical and thermal stability as well as highly sensitive towards both complementary and non-complementary sequences, promoted them in the Olympus of biocomponents for applications in diagnostics. The intent of this review (85 ref) is to describe the last trends on the astonishing PNA technology in the biosensing field, in the view of the positive contamination with vanguard technologies including rational design and nanotechnology. In particular, electrochemical and optical biosensors exploiting PNA were reported from the literature, as well as nanostructured and paper-based biosensors for the design of microfluidics and lab-on-chip tools. • PNAs mimic DNA in binding oligonucleotide sequences through Hoogsteen base pairing in the major groove. • PNAs show outstanding stability and sensitivity towards both complementary and non-complementary sequences. • Vanguard technologies as rational design, nanotechnology, and microfluidics boost PNA use as smart bioreceptors for biosing. [ABSTRACT FROM AUTHOR]