原核Argonaute的应用研究进展

原核生物Argonautes （pAgos）是参与细胞防御外来DNA入侵的可编程核酸酶。在体外，pAgos可以结合小的单链核酸（ssDNA/ssRNA）向导来识别和切割互补DNA/RNA。在体内，pAgos优先靶向多拷贝遗传元件、噬菌体和质粒，从而抑制入侵核酸的扩增和噬菌体感染。pAgos作为一类新兴的可编程核酸酶，比目前应用最为广泛的CRISPR-Cas系统更具灵活性，在生物技术方面展现出巨大的潜力。早期的研究聚焦于嗜热的pAgo，目前基于嗜热pAgos的主要应用包括分子诊断和体外DNA组装。为了推进基于Ago的体内生物技术，如基因编辑的应用，研究人员的焦点逐渐转移到中温生物来源的pAgos，虽然目前pAgos还未实现基因组编辑，但是随着越来越多的pAgo被发掘以及研究人员对pAgos催化机制的深入研究，有望开发基于pAgos的下一代基因编辑技术。本文总结了已知代表性pAgos和基于pAgos发展的生物技术，并简要分析了pAgos在原核生物和真核生物体内应用面临的挑战和可能的应对策略。

Progress on application of prokaryotic Argonaute

Prokaryotic Argonautes （pAgos） are programmable nucleases involved in cellular defense against foreign DNA invasion. In vitro， pAgos can bind to small single stranded guide nucleic acid （ssDNA/ssRNA） to recognize and cleave complementary DNA/RNA. In vivo， pAgos preferentially target multiple copies of genetic elements， bacteriophages， and plasmids， thereby inhibiting the propagation of invading nucleic acids and bacteriophage infections. Prokaryotic Argonautes （pAgos）， as an emerging class of programmable nucleases， are more flexible than the most widely used CRISPR Cas system and has shown great potential in biotechnology. Previous studies were primarily focused on the thermophilic pAgos. Nowadays， the main applications based on thermophilic pAgos include molecular diagnosis and DNA assembly in vitro. Researchers have gradually shifted their focus to pAgos from mesophilic biological sources to promote the application of Ago-based biotechnology in vivo， such as gene editing. Although genome editing has not yet been achieved by pAgos， it’s possible to develop the next generation techniques for genome-editing based on pAgos with more and more pAgos being discovered and researchers studying the catalytic mechanism of pAgos in depth. This article summarized the known representative pAgos and biotechnologies based on the development of pAgos. The challenges and potential strategies faced by the application of pAgos in prokaryotic and eukaryotic organisms were briefly analyzed.