Fig. 6: Cas13d antivirals can treat established viral infection in human primary lung epithelial cultures.
From: Broad-spectrum CRISPR-mediated inhibition of SARS-CoV-2 variants and endemic coronaviruses in vitro
a Scheme used for antiviral treatment of air–liquid-interface (ALI) cultures of human primary bronchial epithelial cells (hPBECs). hPBECs were transduced with NES-Cas13d and differentiated to ALI cultures. The ALI cultures were infected with 229E at an MOI of 0.05, with SARS-CoV-2 WA1 at an MOI of 0.6, or with Omicron at an MOI of 0.1. At 6 hpi, the culture was transfected with crRNA using LNP. At 48, 72 hpi, the apical surfaces of ALI cultures were washed with PBS and the wash solution was collected for measuring virus titer. b The titer of 229E virus released on the apical surface of ALI cultures was determined by RT-qPCR; n = 4, t = 3. c The titer of SARS-CoV-2 virus, including WA1 and Omicron strains, was determined at 48 hpi; n = 3, t = 3. n is the number of independent biological experiments. t is the number of technician replicates per biological replicate in the RT-qPCR assay. All source data in this figure are provided as a Source data file. P values are listed in supplementary Data 3, calculated by two-tailed Student’s t test.
