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

Hyunjae Chung; Maxwell P Bui-Marinos; Waleed Rahmani; Jennifer A Corcoran; Justin Chun

doi:10.1016/j.xpro.2022.101617

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

STAR Protoc. 2022 Jul 21;3(3):101617. doi: 10.1016/j.xpro.2022.101617. eCollection 2022 Sep 16.

Authors

Hyunjae Chung¹, Maxwell P Bui-Marinos², Waleed Rahmani³, Jennifer A Corcoran⁴, Justin Chun⁵

Affiliations

¹ Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
² Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada; Microbiology, Immunology and Infectious Diseases Department and Charbonneau Cancer Research Institute, University of Calgary, Calgary, AB, Canada.
³ Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
⁴ Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada; Microbiology, Immunology and Infectious Diseases Department and Charbonneau Cancer Research Institute, University of Calgary, Calgary, AB, Canada. Electronic address: jennifer.corcoran@ucalgary.ca.
⁵ Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada; Department of Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada. Electronic address: chuj@ucalgary.ca.

Abstract

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).

Keywords: Cell-based assays; Organoids; Stem cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19*
Clone Cells
DNA, Complementary / genetics
Humans
Kidney
Organoids
SARS-CoV-2* / genetics

Substances

DNA, Complementary

Grants and funding

175622/CIHR/Canada