A CRISPR/Cas9 genetically engineered organoid biobank reveals essential host factors for coronaviruses

Autor: Jelte van der Vaart, Bart L. Haagmans, Anna Z Mykytyn, Georg A. Busslinger, Tim I Breugem, Mart M. Lamers, Maarten H. Geurts, Wim de Lau, Cayetano Pleguezuelos-Manzano, Joep Beumer, Jingshu Zhang, Debby Schipper, Petra B. van den Doel, Hans Clevers, Jens Puschhof, Samra Riesebosch
Přispěvatelé: Virology, Hubrecht Institute for Developmental Biology and Stem Cell Research
Jazyk: angličtina
Rok vydání: 2021
Předmět:
CRISPR-Cas9 genome editing
Proteases
Middle East respiratory syndrome coronavirus
Science
Dipeptidyl Peptidase 4
viruses
Mutant
General Physics and Astronomy
Serine Endopeptidases/genetics
Computational biology
Biology
medicine.disease_cause
Virus Replication
General Biochemistry
Genetics and Molecular Biology
Article
Stem-cell biotechnology
Cell Line
Dipeptidyl Peptidase 4/genetics
SDG 3 - Good Health and Well-being
medicine
Organoid
Organoids/metabolism
CRISPR
Humans
Gene
Coronavirus
Biological Specimen Banks
Multidisciplinary
SARS-CoV-2
Intestinal stem cells
Serine Endopeptidases
virus diseases
COVID-19
General Chemistry
Organoids
Angiotensin-Converting Enzyme 2/genetics
Viral replication
Genetic engineering
Middle East Respiratory Syndrome Coronavirus
Angiotensin-Converting Enzyme 2
CRISPR-Cas Systems
Transcriptome
Genetic screen
Zdroj: Nature Communications
Nature Communications, 12(1):5498. Nature Publishing Group
Nature Communications, Vol 12, Iss 1, Pp 1-12 (2021)
Nature Communications, 12(1). Nature Publishing Group
ISSN: 2041-1723
Popis: 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 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.
Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Here the authors generate mutant clonal intestinal organoids for 19 host genes previously implicated in coronavirus biology and identify the cell surface protease TMPRSS2 as a potential therapeutic target.
Databáze: OpenAIRE
Externí odkaz: