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Title: Effective suppression of yellow head virus replication in Penaeus monodon hemocytes using constitutive expression vector for long-hairpin RNA (lhRNA)
Authors: Phathrawarin Thedcharoen
Yongyut Pewkliang
Hoa Khanh Tran Kiem
Lalana Nuntakarn
Suparat Taengchaiyaphum
Kallaya Sritunyalucksana
Timothy W. Flegel
Vanvimon Saksmerprome
Suparerk Borwornpinyo
Mahidol University
Thailand National Center for Genetic Engineering and Biotechnology
Keywords: Agricultural and Biological Sciences
Issue Date: 1-Sep-2020
Citation: Journal of Invertebrate Pathology. Vol.175, (2020)
Abstract: © 2020 Elsevier Inc. Double-stranded RNA (dsRNA) is employed to down-regulate the expression of specific genes of shrimp viral pathogens through the RNA interference (RNAi) pathway. The administration of dsRNA into shrimp has been shown to be an effective strategy to block yellow head virus (YHV) progression. In this study, a vector (pLVX-AcGFP1-N1) was developed to introduce a long-hairpin RNA (lhRNA) silencing cassette under a CMV promoter, so-called “pLVX-lhRdRp”, against the RNA-dependent RNA polymerase (RdRp) gene of YHV. A primary culture of hemocytes isolated from Penaeus monodon was transfected with the pLVX-lhRdRp vector, generating transcripts of lhRNAs as early as 12 h post transfection. Twelve hours prior to YHV challenge, the primary hemocyte cell culture was transfected with pLVX-lhRdRp, whereas control groups were transfected with pLVX-AcGFP1-N1 or no transfection. The group treated with pLVX-lhRdRp significantly suppressed YHV replication at 24–72 h after YHV challenge. The results from RT-PCR and immunohistochemistry confirmed that both mRNA and protein expression of YHV were effectively inhibited by the pLVX-lhRdRp vector. Thus, our hemocyte culture and dsRNA expression plasmid with constitutive promoter have potential as a platform to test DNA constructs expressing long-hairpin RNA against pathogenic viral infection and as a RNAi-based DNA vaccine in shrimp.
ISSN: 10960805
Appears in Collections:Scopus 2020

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