Aerolysin is one of the most studied beta pore-forming toxins and has recently emerged as a powerful tool in nanopore sensing due to its narrow and charged pore lumen. By monitoring ionic current changes as single molecules pass through the pore, aerolysin enables the detection of a wide variety of biomolecules with high sensitivity. In our lab, we have demonstrated aerolysin’s ability to discriminate single nucleotides and peptides and to detect post-translational modifications used as biomarkers for neurodegenerative diseases such as Parkinson’s disease. Additionally, aerolysin pores have shown potential for decoding digital information stored in synthetic polymers, offering a novel approach to address challenges in data storage. Despite its diverse applications, the atomic-resolution structure of aerolysin had remained elusive. We recently published the first high-resolution cryo-EM structure of aerolysin in several membrane mimics including copolymer nanodiscs. Using this membrane-like environment allowed the identification of lipids surrounding the pore and to determine its correct position in the membrane. Moreover, the structure revealed key interactions necessary for membrane anchoring and allowed to better understand the pore formation mechanism. Notably, we were also able to identify four constriction rings in the pore lumen which are highly relevant for nanopore sensing. The high-resolution structures will allow for more precise modeling and engineering of aerolysin in the future. As a next step, we aim to discover currently unknown PFTs that could allow completely new sensing applications. By searching the AlphaFold database we identified several promising candidates that we are currently characterizing biophysically and in in the nanopore setup.