tetano
Editor, Senior Moderator
Cell Rep Methods
. 2022 Feb 1;100170.
doi: 10.1016/j.crmeth.2022.100170. Online ahead of print.
Multi-color super-resolution imaging to study human coronavirus RNA during cellular infection
Jiarui Wang[SUP] 1 2 [/SUP], Mengting Han[SUP] 3 [/SUP], Anish R Roy[SUP] 1 [/SUP], Haifeng Wang[SUP] 3 [/SUP], Leonhard Möckl[SUP] 1 [/SUP], Leiping Zeng[SUP] 3 [/SUP], W E Moerner[SUP] 1 [/SUP], Lei S Qi[SUP] 3 [/SUP]
Affiliations
Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the third human coronavirus within 20 years that gave rise to a life-threatening disease and the first to reach pandemic spread. To make therapeutic headway against current and future coronaviruses, the biology of coronavirus RNA during infection must be precisely understood. Here, we present a robust and generalizable framework combining high-throughput confocal and super-resolution microscopy imaging to study coronavirus infection at the nanoscale. Employing the model human coronavirus HCoV-229E, we specifically labeled coronavirus genomic RNA (gRNA) and double-stranded RNA (dsRNA) via multicolor RNA-immunoFISH and visualized their localization patterns within the cell. The 20nm resolution achieved by of our approach uncovers a striking spatial organization of gRNA and dsRNA into three distinct structures and enables quantitative characterization of the status of the infection after antiviral drug treatment. Our approach provides a comprehensive imaging framework that will enable future investigations of coronavirus fundamental biology and therapeutic effects.
Keywords: ER remodeling; FISH; RNA localization; Remdesivir; coronavirus infection model; fluorescence imaging; human coronavirus 229E; spike protein; super-resolution microscopy; viral replication and infection.
. 2022 Feb 1;100170.
doi: 10.1016/j.crmeth.2022.100170. Online ahead of print.
Multi-color super-resolution imaging to study human coronavirus RNA during cellular infection
Jiarui Wang[SUP] 1 2 [/SUP], Mengting Han[SUP] 3 [/SUP], Anish R Roy[SUP] 1 [/SUP], Haifeng Wang[SUP] 3 [/SUP], Leonhard Möckl[SUP] 1 [/SUP], Leiping Zeng[SUP] 3 [/SUP], W E Moerner[SUP] 1 [/SUP], Lei S Qi[SUP] 3 [/SUP]
Affiliations
- PMID: 35128513
- PMCID: PMC8806145
- DOI: 10.1016/j.crmeth.2022.100170
Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the third human coronavirus within 20 years that gave rise to a life-threatening disease and the first to reach pandemic spread. To make therapeutic headway against current and future coronaviruses, the biology of coronavirus RNA during infection must be precisely understood. Here, we present a robust and generalizable framework combining high-throughput confocal and super-resolution microscopy imaging to study coronavirus infection at the nanoscale. Employing the model human coronavirus HCoV-229E, we specifically labeled coronavirus genomic RNA (gRNA) and double-stranded RNA (dsRNA) via multicolor RNA-immunoFISH and visualized their localization patterns within the cell. The 20nm resolution achieved by of our approach uncovers a striking spatial organization of gRNA and dsRNA into three distinct structures and enables quantitative characterization of the status of the infection after antiviral drug treatment. Our approach provides a comprehensive imaging framework that will enable future investigations of coronavirus fundamental biology and therapeutic effects.
Keywords: ER remodeling; FISH; RNA localization; Remdesivir; coronavirus infection model; fluorescence imaging; human coronavirus 229E; spike protein; super-resolution microscopy; viral replication and infection.