About: Establishment of primary transgenic human airway epithelial cell cultures to study respiratory virus

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About: Establishment of primary transgenic human airway epithelial cell cultures to study respiratory virus – host interactions Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Establishment of primary transgenic human airway epithelial cell cultures to study respiratory virus – host interactions
Creator	Thiel, Volker Dijkman, Ronald Jonsdottir, Hulda Rodriguez, Regulo Geerts, Dirk »more»
source	BioRxiv; MedRxiv
abstract	Primary human airway epithelial cell (hAEC) cultures represent a universal platform to propagate respiratory viruses and characterize their host interactions in authentic target cells. To further elucidate specific interactions between human respiratory viruses and important host factors in airway epithelium, it is important to make hAEC cultures amenable to genetic modification. However, the short and finite lifespan of primary cells in cell culture creates a bottleneck for the genetic modification of these cultures. In the current study, we show that the incorporation of the Rho-associated protein kinase (ROCK) inhibitor (Y-27632) during cell propagation extends the life span of primary human cells in vitro and thereby facilitates the incorporation of lentivirus-based expression systems. Using fluorescent reporters for FACS-based sorting, we generated homogenously fluorescent hAEC cultures that differentiate normally after lentiviral transduction. As proof-of-principle, we demonstrate that host gene expression can be modulated post-differentiation via inducible short hairpin (sh)RNA-mediated knockdown. Importantly, functional characterization of these transgenic hAEC cultures with exogenous poly(I:C), as a proxy for virus infection, demonstrates that such modifications do not influence the host innate immune response. Moreover, the propagation kinetics of both human coronavirus 229E (HCoV-229E) and human respiratory syncytial virus (RSV) were not affected. Combined, these results validate our newly established protocol for the genetic modification of hAEC cultures thereby unlocking a unique potential for detailed molecular characterization of virus – host interactions in human respiratory epithelium.
has issue date	2019-07-08(xsd:dateTime)
bibo:doi	10.1101/694380
has license	medrxiv
sha1sum (hex)	891117a658301b282d3f889711b467e832b68cb1
schema:url	https://doi.org/10.1101/694380
resource representing a document's title	Establishment of primary transgenic human airway epithelial cell cultures to study respiratory virus – host interactions
schema:publication	bioRxiv
resource representing a document's body	covid:891117a658301b282d3f889711b467e832b68cb1#body_text
is schema:about of	named entity 'viruses' named entity 'platform' named entity 'human' named entity 'host' named entity 'propagate' »more»

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