About: Recombinant Pseudorabies Virus (PRV) Expressing Firefly Luciferase Effectively Screened for CRISPR/Cas9 Single Guide RNAs and Antiviral Compounds

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About: Recombinant Pseudorabies Virus (PRV) Expressing Firefly Luciferase Effectively Screened for CRISPR/Cas9 Single Guide RNAs and Antiviral Compounds 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	Recombinant Pseudorabies Virus (PRV) Expressing Firefly Luciferase Effectively Screened for CRISPR/Cas9 Single Guide RNAs and Antiviral Compounds
Creator	An, Tong-Qing Tian, Zhi-Jun Cai, Xue-Hui Tang, Yan-Dong Fang, Qiong-Qiong Wang, Tong-Yun Sun, Ming-Xia Liu, Ji-Ting
Source	Medline; PMC
abstract	A Pseudorabies virus (PRV) variant has emerged in China since 2011 that is not protected by commercial vaccines, and has not been well studied. The PRV genome is large and difficult to manipulate, but it is feasible to use clustered, regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology. However, identification of single guide RNA (sgRNA) through screening is critical to the CRISPR/Cas9 system, and is traditionally time and labor intensive, and not suitable for rapid and high throughput screening of effective PRV sgRNAs. In this study, we developed a recombinant PRV strain expressing firefly luciferase and enhanced green fluorescent protein (EGFP) as a reporter virus for PRV-specific sgRNA screens and rapid evaluation of antiviral compounds. Luciferase activity was apparent as soon as 4 h after infection and was stably expressed through 10 passages. In a proof of the principle screen, we were able to identify several PRV specific sgRNAs and confirmed that they inhibited PRV replication using traditional methods. Using the reporter virus, we also identified PRV variants lacking US3, US2, and US9 gene function, and showed anti-PRV activity for chloroquine. Our results suggest that the reporter PRV strain will be a useful tool for basic virology studies, and for developing PRV control and prevention measures.
has issue date	2016-03-29(xsd:dateTime)
bibo:doi	10.3390/v8040090
bibo:pmid	27043610
has license	cc-by
sha1sum (hex)	5dd71c3b90ad6f642495062f9f591f78fccd8d75
schema:url	https://doi.org/10.3390/v8040090
resource representing a document's title	Recombinant Pseudorabies Virus (PRV) Expressing Firefly Luciferase Effectively Screened for CRISPR/Cas9 Single Guide RNAs and Antiviral Compounds
has PubMed Central identifier	PMC4848585
has PubMed identifier	27043610
schema:publication	Viruses
resource representing a document's body	covid:5dd71c3b90ad6f642495062f9f591f78fccd8d75#body_text
is schema:about of	named entity 'SUGGEST' named entity 'EVALUATION' named entity 'LABOR' named entity 'BASIC' named entity 'ANTIVIRAL' named entity 'APPARENT' named entity 'SPECIFIC' named entity 'STRAIN' named entity 'TIME' named entity 'BUT' named entity 'US2' named entity 'LACKING' named entity 'US9' named entity 'METHODS' named entity 'CHLOROQUINE' named entity 'IDENTIFICATION' named entity 'PREVENTION MEASURES' named entity 'SCREENING' named entity 'CRISPR' named entity 'homologous arms' named entity 'hpi' named entity 'dsDNA' named entity 'virus' named entity 'Hepatitis C Virus' named entity 'sequence data' named entity 'viral protein' named entity 'Vero cells' named entity 'protease inhibitor' named entity 'base pairs' named entity 'error-prone' named entity 'CsA' named entity 'HSV-1' named entity 'Cas9' named entity 'CsA' named entity 'melting point' named entity 'antiviral' named entity 'homologous arms' named entity 'Vero cells' named entity 'Cas9' named entity 'US3' named entity 'US3' named entity 'vector' named entity 'culture media' named entity 'Clontech' named entity 'plasmid' named entity 'SpCas9' named entity 'randomly selected' named entity 'PRV' named entity 'CMV' named entity 'Cas9' named entity 'PRRSV' named entity 'sequenced' named entity 'cas9' named entity 'EGFP' named entity 'Switzerland' named entity 'DNA viruses' named entity 'Cas9' named entity 'antiviral' named entity 'zinc-finger nucleases' named entity 'sgRNA' named entity 'PaloAlto' named entity 'TALENs' named entity 'SV40' named entity 'EGFP' named entity 'viral replication' named entity 'PRV' named entity 'luciferase' named entity 'primers' named entity 'SDS-PAGE' named entity 'transfected' named entity 'virus' named entity 'Thermo scientific' named entity 'gene' named entity 'ZFNs'

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