Rapid identification of host
genes essential for
virus replication may expedite the generation of
therapeutic interventions. Genetic
screens are often performed in
transformed cell lines that poorly represent viral target
cells in vivo, leading to discoveries that may not be translated to the clinic. Intestinal
organoids (IOs) are increasingly used to model
human disease and are amenable to
genetic engineering. To discern which host factors are reliable anti-
coronavirus therapeutic targets, we generate mutant clonal IOs for 19 host
genes previously implicated in
coronavirus biology. We verify ACE2 and DPP4 as entry receptors for
SARS-CoV/
SARS-CoV-2 and
MERS-CoV respectively.
SARS-CoV-2 replication in IOs does not require the endosomal
Cathepsin B/L
proteases, but specifically depends on the
cell surface
protease TMPRSS2. Other TMPRSS
family members were not essential. The newly emerging
coronavirus variant B.1.1.7, as well as
SARS-CoV and
MERS-CoV similarly depended on TMPRSS2. These findings underscore the relevance of non-transformed
human models for
coronavirus research, identify TMPRSS2 as an attractive pan-
coronavirus therapeutic target, and demonstrate that an
organoid knockout biobank is a valuable tool to investigate the
biology of current and
future emerging
coronaviruses.