دورية أكاديمية
Effective suppression of yellow head virus replication in Penaeus monodon hemocytes using constitutive expression vector for long-hairpin RNA (lhRNA).
| العنوان: | Effective suppression of yellow head virus replication in Penaeus monodon hemocytes using constitutive expression vector for long-hairpin RNA (lhRNA). |
|---|---|
| المؤلفون: | Thedcharoen P; Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand. Electronic address: phathrawarin.t@gmail.com., Pewkliang Y; Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand; Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok, Thailand. Electronic address: yongyut.pew@gmail.com., Kiem HKT; Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand. Electronic address: tkhoa21@yahoo.com., Nuntakarn L; Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand. Electronic address: lalana.nun@gmail.com., Taengchaiyaphum S; Aquatic Animal Health Research Team (AQHT), Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok, Thailand. Electronic address: suparatmb@gmail.com., Sritunyalucksana K; Aquatic Animal Health Research Team (AQHT), Integrative Aquaculture Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok, Thailand. Electronic address: kallaya@biotec.or.th., Flegel TW; Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand; Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok, Thailand; National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand. Electronic address: timothy.fle@mahidol.ac.th., Saksmerprome V; Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok, Thailand; National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand. Electronic address: vanvimon.sak@biotec.or.th., Borwornpinyo S; Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand; Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok, Thailand; Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok, Thailand. Electronic address: suparerk.bor@mahidol.ac.th. |
| المصدر: | Journal of invertebrate pathology [J Invertebr Pathol] 2020 Sep; Vol. 175, pp. 107442. Date of Electronic Publication: 2020 Jul 12. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 0014067 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0805 (Electronic) Linking ISSN: 00222011 NLM ISO Abbreviation: J Invertebr Pathol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: New York, NY : Academic Press Original Publication: New York. |
| مواضيع طبية MeSH: | RNA Interference* , Virus Replication*, Hemocytes/*virology , Penaeidae/*virology , RNA, Double-Stranded/*metabolism , Roniviridae/*physiology, Animals |
| مستخلص: | 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. Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2020 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Penaeus monodon shrimp; Primary hemocyte culture; RNAi; Yellow head virus (YHV); dsRNA; lhRNA |
| المشرفين على المادة: | 0 (RNA, Double-Stranded) |
| تواريخ الأحداث: | Date Created: 20200715 Date Completed: 20210715 Latest Revision: 20210715 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.jip.2020.107442 |
| PMID: | 32663545 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1096-0805 |
|---|---|
| DOI: | 10.1016/j.jip.2020.107442 |