About: A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination

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About: A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination Goto Sponge NotDistinct Permalink

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

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination
Creator	Rottier, Peter Krämer-Kühl, Annika Mundt, Egbert Chénard, Gilles Philipp, Hans-Christian Spekreijse, Dieuwertje Berends, Alinda Hélène Verheije, M Van Beurden, Steven
Source	Medline; PMC
abstract	BACKGROUND: Avian coronavirus infectious bronchitis virus (IBV) is a respiratory pathogen of chickens that causes severe economic losses in the poultry industry worldwide. Major advances in the study of the molecular biology of IBV have resulted from the development of reverse genetics systems for the highly attenuated, cell culture-adapted, IBV strain Beaudette. However, most IBV strains, amongst them virulent field isolates, can only be propagated in embryonated chicken eggs, and not in continuous cell lines. METHODS: We established a reverse genetics system for the IBV strain H52, based on targeted RNA recombination in a two-step process. First, a genomic and a chimeric synthetic, modified IBV RNA were co-transfected into non-susceptible cells to generate a recombinant chimeric murinized (m) IBV intermediate (mIBV). Herein, the genomic part coding for the spike glycoprotein ectodomain was replaced by that of the coronavirus mouse hepatitis virus (MHV), allowing for the selection and propagation of recombinant mIBV in murine cells. In the second step, mIBV was used as the recipient. To this end a recombination with synthetic RNA comprising the 3′-end of the IBV genome was performed by introducing the complete IBV spike gene, allowing for the rescue and selection of candidate recombinants in embryonated chicken eggs. RESULTS: Targeted RNA recombination allowed for the modification of the 3′-end of the IBV genome, encoding all structural and accessory genes. A wild-type recombinant IBV was constructed, containing several synonymous marker mutations. The in ovo growth kinetics and in vivo characteristics of the recombinant virus were similar to those of the parental IBV strain H52. CONCLUSIONS: Targeted RNA recombination allows for the generation of recombinant IBV strains that are not able to infect and propagate in continuous cell lines. The ability to introduce specific mutations holds promise for the development of rationally designed live-attenuated IBV vaccines and for studies into the biology of IBV in general. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12985-017-0775-8) contains supplementary material, which is available to authorized users.
has issue date	2017-06-12(xsd:dateTime)
bibo:doi	10.1186/s12985-017-0775-8
bibo:pmid	28606144
has license	cc-by
sha1sum (hex)	a9cab864aceb9a873c6a049a8d7951ac80d993d8
schema:url	https://doi.org/10.1186/s12985-017-0775-8
resource representing a document's title	A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination
has PubMed Central identifier	PMC5468965
has PubMed identifier	28606144
schema:publication	Virol J
resource representing a document's body	covid:a9cab864aceb9a873c6a049a8d7951ac80d993d8#body_text
is schema:about of	named entity 'process' named entity 'mouse' named entity 'development' named entity 'study' named entity 'modified' named entity 'However' named entity 'Avian' named entity 'TARGETED' named entity 'METHODS' named entity 'RNA RECOMBINATION' named entity 'COMPLETE' named entity 'SELECTION' covid:arg/a9cab864aceb9a873c6a049a8d7951ac80d993d8 named entity 'PROCESS' named entity 'PERFORMED BY' named entity 'ISOLATES' named entity 'MURINE' named entity 'RECIPIENT' named entity 'TARGETED' named entity 'CAUSES' named entity 'IS A' named entity 'SYNTHETIC' named entity 'GENOME' named entity 'NONSUSCEPTIBLE' named entity 'FIELD' named entity 'STRAIN' named entity 'CANDIDATE' named entity 'BASED' named entity 'INFECTIOUS BRONCHITIS VIRUS' named entity 'AVIAN CORONAVIRUS' named entity 'REVERSE GENETICS' named entity 'H52' named entity 'AVIAN CORONAVIRUS' named entity 'GENE' named entity 'RNA' named entity 'CHIMERIC' named entity 'CONTINUOUS' named entity 'HIGHLY' named entity 'SECOND' named entity 'STEP' named entity 'SYSTEM' named entity 'CORONAVIRUS' named entity 'CHICKEN' named entity 'REVERSE GENETICS' named entity 'RECOMBINANT' named entity 'ECTODOMAIN' named entity 'BACKGROUND' named entity 'ECONOMIC' named entity 'HAVE' named entity 'VIRULENT' named entity '80%' named entity 'STUDY' named entity 'EGGS' named entity 'DEVELOPMENT' named entity 'GENOMIC' named entity 'BASED' named entity 'CELL CULTURE' named entity 'SEVERE' named entity 'RESPIRATORY' named entity 'RECOMBINATION' named entity 'CHICKENS' named entity 'CODING' named entity 'END' named entity 'POULTRY INDUSTRY' named entity 'MOUSE HEPATITIS VIRUS' named entity 'STRAINS' named entity 'REPLACED BY' named entity 'RECOMBINANTS' named entity 'SYSTEMS' named entity 'PATHOGEN' named entity 'WORLDWIDE' named entity 'ESTABLISHED' named entity 'SYSTEM'

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