New RNA aptamer-dye systems for MINFLUX, super-resolution, and single-molecule imaging

• Applicant: Professor Dr. Andres Jäschke
• Subject Area: Biological and Biomimetic Chemistry
• Term: since 2017
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 339113386

Fluorescent RNA imaging has long been challenged by limitations in sensitivity and resolution, restricting our ability to study RNA dynamics within living cells. Given the pivotal role of RNA in cell biology, it is imperative to develop precise and sensitive probes and techniques for real-time RNA imaging in vivo. Our research group has made significant strides in advancing RNA imaging technologies, particularly with the introduction of the first fluorescent light-up aptamers (FLAPs) enabling super-resolution imaging through techniques like SMLM and STED. The field of super-resolution microscopy has seen rapid growth, fueled by innovations such as the MINFLUX technique and notable improvements in PAINT methods. Building upon our previous accomplishments, our research project outlines five key objectives: i) Develop FLAP-dye systems that push the boundaries of biological RNA imaging to attain single-digit nanometer resolution. ii) Broaden the utility of FLAPs by creating super-resolution-capable variants tailored for different spectral ranges, with a specific focus on near-infrared and green wavelengths. iii) Explore additional physical parameters, such as fluorescence lifetime, to further enhance FLAP-based RNA imaging techniques. iv) Apply the spirolactonization concept to avidity probes, thereby elevating their performance and versatility. v) Extend the capabilities of our high-performance FLAPs to encompass the imaging of proteins both within and on cells and on tissue sections. These objectives represent an ambitious yet essential next step in our efforts to revolutionize the field of RNA and protein imaging, ultimately advancing our understanding of fundamental cellular processes.

• DFG Programme: Research Grants