Prokaryotic Argonaute Protein from Natronobacterium gregoryi Requires RNAs To Activate for DNA Interference
            <i>In Vivo</i>

Xing, Jiani; Ma, Lei; Cheng, Xinzhen; Ma, Jinrong; Wang, Ruyu; Xu, Kun; Mymryk, Joe S.; Zhang, Zhiying

doi:10.1128/mbio.03656-21

Cited by 3 publications

(1 citation statement)

References 30 publications

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“…Eukaryotic Argonaute proteins (eAgos) are a core of eukaryotic RNA interference (RNAi) machinery [ 1 , 2 ], which use RNA guides to recognize and cleave complementary RNA targets. Prokaryotic Argonautes (pAgos) are found in bacteria and archaea [ 3 , 4 , 5 , 6 ], such as TtAgo from Thermus thermophilus [ 7 ], PfAgo from Pyrococcus furiosus [ 8 ], and MjAgo from Methanocaldococcus jannaschii [ 9 ]. These pAgos prefer to cleave DNA in vitro and protect cells from foreign genetic elements in vivo [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

A Programmable, DNA-Exclusively-Guided Argonaute DNase and Its Higher Cleavage Specificity Achieved by 5′-Hydroxylated Guide

Sun

Zhu

et al. 2022

Biomolecules

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Argonaute proteins exist widely in eukaryotes and prokaryotes, and they are of great potential for molecular cloning, nucleic acid detection, DNA assembly, and gene editing. However, their overall properties are not satisfactory and hinder their broad applications. Herein, we investigated a prokaryotic Argonaute nuclease from a mesophilic bacterium Clostridium disporicum (CdAgo) and explored its overall properties, especially with 5’-hydroxylated (5’-OH) guides. We found that CdAgo can exclusively use single-stranded DNA (ssDNA) as guide to cleave ssDNA and plasmid targets. Further, we found the length of the efficient guide is narrower for the 5’-OH guide (17–20 nt) than for the 5′-phosphorylated guide (5′-P, 14–21 nt). Furthermore, we discovered that the 5’-OH guides can generally offer stronger mismatch discrimination than the 5′-P ones. The 5’-OH guides offer the narrower length range, higher mismatch discrimination and more accurate cleavage than the 5′-P guides. Therefore, 5’-OH-guide-directed CdAgo has great potential in biological and biomedical applications.

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