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Cryo-EM and antisense targeting of the 28-kDa frameshift stimulation element from the SARS-CoV-2 RNA genome
Nature Structural & Molecular Biology ( IF 12.5 ) Pub Date : 2021-08-23 , DOI: 10.1038/s41594-021-00653-y
Kaiming Zhang 1, 2 , Ivan N Zheludev 3 , Rachel J Hagey 4 , Raphael Haslecker 5 , Yixuan J Hou 6 , Rachael Kretsch 7 , Grigore D Pintilie 1 , Ramya Rangan 7 , Wipapat Kladwang 3 , Shanshan Li 1, 2 , Marie Teng-Pei Wu 5 , Edward A Pham 4 , Claire Bernardin-Souibgui 4 , Ralph S Baric 6, 8 , Timothy P Sheahan 6 , Victoria D'Souza 5 , Jeffrey S Glenn 4, 9 , Wah Chiu 1, 7, 10 , Rhiju Das 3, 7, 11
Affiliation  

Drug discovery campaigns against COVID-19 are beginning to target the SARS-CoV-2 RNA genome. The highly conserved frameshift stimulation element (FSE), required for balanced expression of viral proteins, is a particularly attractive SARS-CoV-2 RNA target. Here we present a 6.9 Å resolution cryo-EM structure of the FSE (88 nucleotides, ~28 kDa), validated through an RNA nanostructure tagging method. The tertiary structure presents a topologically complex fold in which the 5′ end is threaded through a ring formed inside a three-stem pseudoknot. Guided by this structure, we develop antisense oligonucleotides that impair FSE function in frameshifting assays and knock down SARS-CoV-2 virus replication in A549-ACE2 cells at 100 nM concentration.



中文翻译:


SARS-CoV-2 RNA 基因组中 28 kDa 移码刺激元件的冷冻电镜和反义靶向



针对 COVID-19 的药物发现活动开始针对 SARS-CoV-2 RNA 基因组。病毒蛋白平衡表达所需的高度保守的移码刺激元件 (FSE) 是一个特别有吸引力的 SARS-CoV-2 RNA 靶点。在这里,我们展示了 FSE 的 6.9 Å 分辨率冷冻电镜结构(88 个核苷酸,约 28 kDa),并通过 RNA 纳米结构标记方法进行了验证。三级结构呈现出拓扑复杂的折叠,其中 5' 端穿过三茎假结内形成的环。在这种结构的指导下,我们开发了反义寡核苷酸,可在移码测定中损害 FSE 功能,并以 100 nM 浓度抑制 A549-ACE2 细胞中 SARS-CoV-2 病毒的复制。

更新日期:2021-08-23
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