Functional Aptamers\nIn Vitro Evolution for Intranuclear\nBlockage of RNA-Protein Interaction

Over the last 30 years, despite considerable research\nand endeavors\naimed at harnessing aptamers as pharmaceutical molecules, the progress\nin developing aptamer-based drugs has been falling short of expectations.\nSequential steps of affinity molecule acquisition and functional screening\nare typically required for discovering affinity-based macromolecule\ntherapeutics, which can be time-consuming and limiting in candidate\nselection. Additionally, aptamers often necessitate tedious postselection\nmodifications to overcome pharmacokinetic limitations, which usually\nimpede the binding affinity. Herein, we propose a novel in vitro screening\nplatform termed Functional Aptamers in vitro Evolution (FAIVE), which\nintegrates affinity molecule acquisition with functional screening\nand introduces chemical diversity during the process. This platform\naims to rapidly generate functional aptamers capable of binding to\ntarget proteins and regulating their functions. Illustrated by targeting\nintranuclear RNA-protein interactions involving HIV-1 Tat protein\nand TAR RNA, FAIVE demonstrates a selection of functional aptamers\nwith significant intracellular blocking effects. The study also explores\nlipid nanoparticle delivery systems to enhance intracellular delivery\nefficiency, expanding aptamer targeting potential to broader intracellular\nand intranuclear domains. This study emphasizes the potential of FAIVE\nto expedite the development of aptamer-based drugs and facilitate\nthe creation of more versatile and effective therapeutics.