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

STAR protocols (2022-08-23)
Hyunjae Chung, Maxwell P Bui-Marinos, Waleed Rahmani, Jennifer A Corcoran, Justin Chun
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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Fetal Bovine Serum, Canada origin, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Y-27632, InSolution, ≥95%, reversible, inhibitor of Rho kinases