Teams identifies cell entry mechanism of SARS-CoV-2 and therapeutic target for COVID-19 hkbaptistu NatureComms
. Our study reveals that MT1-MMP could be the potential druggable target for these new variants as inhibition of MT1-MMP activity by either pharmacological antagonism or genetic knockdown effectively restricted the cell entry of SARS-CoV-2 wild-type virus and its variants of concern in human primary cells and organoids. Identification of new proteases involved in ACE2 processing will certainly provide potential opportunities to target proteases as an antiviral strategy.
pCMV-GFP-ACE2 and pCMV-FLAG-MMP14 plasmids were purchased from Sino Biological . pCMV-FLAG-MMP14 , the primers used for plasmid construction were as follows: F: GGGGTACCATGTCTCCCGCCCCAAG; R: TCTAGACTCGAGTTTACTTATCGT; F1: TGGTGGCTGTGCACGCGCTGGGCCATGCCCT; R1: AGGGCATGGCCCAGCGCGTGCACAGCCACCA. PCR amplification was performed using the pCMV-FLAG-MMP14 plasmid as a template. The sequences of the gene-specific primers for constructing ACE2 point mutations were shown in Supplementary Table.
. Cardiac organoids were disassembled using TrypLE express upon reaching confluence, and infected with shlentivirus for 24 h. Cardiac organoids were subsequently selected with Puromycin for one week.Cells grown on six-well plates were transfected with 1μg of the expression vector using Lipofectamine 3000 . 48 h after transfection, cells were washed twice with PBS and incubated with serum-free DMEM at 37 °C for 12 h.
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