Advancements of prokaryotic Argonautes in molecular diagnostics and future perspectives

Abstract

Due to the notable biological advantages, programmable nucleases have become a groundbreaking class of tools in the field of nucleic acid testing (NAT). Prokaryotic Argonautes (pAgos), a subclass of Argonaute proteins, represent an emerging class of nucleases that offer greater flexibility compared to conventional CRISPR/Cas systems and hold tremendous biotechnological potential. Recent advancements in pAgo-mediated NAT have opened new avenues for the development of rapid and highly sensitive nucleic acid detection methods. In this review, we first undertake a comprehensive exploration of pAgos in structural diversity and the molecular mechanisms of guide and target binding by pAgos. Further, a comparison analysis of pAgos and CRISPR/Cas nucleases is conducted, focusing on their similarities, differences, and complementarities, with the understanding of what is crucial for the rational design of novel NAT. Afterward, the recent progress of the different pAgo-mediated biosensor platforms is summarized, and the associated challenges and perspectives are discussed. With comprehensive illustration, the goal of this review is to provide new insights and perspectives for developing next-generation nucleic acid detection platforms.