Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2

Induced pluripotent stem cell (iPSC)-derived kidney organoids can be used for disease modeling and drug testing. Here, we describe a protocol to prepare stocks of an infectious clone of SARS-CoV-2 expressing a stable mNeonGreen reporter (icSARS-CoV-2-mNG). We demonstrate the infection of kidney orga...

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Autores principales: Chung, Hyunjae, Bui-Marinos, Maxwell P., Rahmani, Waleed, Corcoran, Jennifer A., Chun, Justin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9300580/
https://www.ncbi.nlm.nih.gov/pubmed/35990736
http://dx.doi.org/10.1016/j.xpro.2022.101617
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author Chung, Hyunjae
Bui-Marinos, Maxwell P.
Rahmani, Waleed
Corcoran, Jennifer A.
Chun, Justin
author_facet Chung, Hyunjae
Bui-Marinos, Maxwell P.
Rahmani, Waleed
Corcoran, Jennifer A.
Chun, Justin
author_sort Chung, Hyunjae
collection PubMed
description Induced pluripotent stem cell (iPSC)-derived kidney organoids can be used for disease modeling and drug testing. Here, we describe a protocol to prepare stocks of an infectious clone of SARS-CoV-2 expressing a stable mNeonGreen reporter (icSARS-CoV-2-mNG). We demonstrate the infection of kidney organoids, primarily at the proximal tubular cells, with icSARS-CoV-2-mNG. Using a TCID50 (tissue culture infectious dose 50) assay and confocal microscopy, we show the quantification of SARS-CoV-2-mNG signal in proximal tubular cells of the kidney organoids. For complete details on the use and execution of this protocol, please refer to Rahmani et al. (2022).
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spelling pubmed-93005802022-07-21 Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2 Chung, Hyunjae Bui-Marinos, Maxwell P. Rahmani, Waleed Corcoran, Jennifer A. Chun, Justin STAR Protoc Protocol Induced pluripotent stem cell (iPSC)-derived kidney organoids can be used for disease modeling and drug testing. Here, we describe a protocol to prepare stocks of an infectious clone of SARS-CoV-2 expressing a stable mNeonGreen reporter (icSARS-CoV-2-mNG). We demonstrate the infection of kidney organoids, primarily at the proximal tubular cells, with icSARS-CoV-2-mNG. Using a TCID50 (tissue culture infectious dose 50) assay and confocal microscopy, we show the quantification of SARS-CoV-2-mNG signal in proximal tubular cells of the kidney organoids. For complete details on the use and execution of this protocol, please refer to Rahmani et al. (2022). Elsevier 2022-07-21 /pmc/articles/PMC9300580/ /pubmed/35990736 http://dx.doi.org/10.1016/j.xpro.2022.101617 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Chung, Hyunjae
Bui-Marinos, Maxwell P.
Rahmani, Waleed
Corcoran, Jennifer A.
Chun, Justin
Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2
title Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2
title_full Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2
title_fullStr Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2
title_full_unstemmed Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2
title_short Infecting kidney organoids with a cDNA reporter clone of SARS-CoV-2
title_sort infecting kidney organoids with a cdna reporter clone of sars-cov-2
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9300580/
https://www.ncbi.nlm.nih.gov/pubmed/35990736
http://dx.doi.org/10.1016/j.xpro.2022.101617
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