RNA interference: the biomolecular approach for in vitro endogenous gene knockdown towards application for prevention of yellow head virus infection in vivo
Event Details
- Date:
- Tuesday, 28 July 2015
- Time:
- 12:00 pm - 1:00 pm
- Room:
- Level 3, Large Seminar Room
- UQ Location:
- Queensland Bioscience Precinct (St Lucia)
- URL:
- http://www.qaafi.uq.edu.au/qaafi-science-seminars-2015#archive
- Event category(s):
Event Contact
- Name:
- Mrs Luba Hickey
- Phone:
- 62092
- Email:
- l.kakoychenko@uq.edu.au
- Org. Unit:
- Queensland Alliance for Agriculture and Food Innovation
Event Description
- Full Description:
- QAAFI Science Seminar
Presented by Assistant Prof. Wanchai Assavalapsakul
Chulalonglkorn University
Thailand
ABSTRACT
The mechanism of RNA interference has been well-recognized as its ability to inhibit gene expression at the transcriptional level through double stranded RNA (dsRNAs) formation. When dsRNAs are formed in cells, they are subject to enzymatic cleavage by Dicer protein to generate small RNAs that subsequently bind to Argonaute protein to form a complex molecule called RNA-Induced Silencing Complex (RISC). RISC inhibits RNA expression by binding to mRNA using the complementary sequence of the small RNA inside the complex to match with the target as well as making use of RNaseIII activity of Argonaute protein, leading to fragments of mRNA and gene silencing. The goal of the present work is to identify specific receptor protein for yellow head virus using RNAi methodology in order to prevent the yellow head infection caused by the virus in black tiger shrimp, Penaeus monodon. In this work, the yellow head virus binding protein (YBP) was characterized by viral overlay protein binding assay and cDNA of YBP was cloned and expressed for the studies, but the YBP sequence was not matched when compared with that in the GenBank database. Therefore, the RNAi technique was introduced to verify whether YBP is a Penaeus monodon yellow head virus receptor protein 65 (PmYRP65). Apparently, after the RNAi knockdown PmYRP65 was implemented in vitro, the viral entry into the cell was forbidden. In addition, silencing endogenous PmYRP65 showed significant survival of the shrimp after infected by the yellow head virus, where approximately 90% of the shrimp could not be infected by the virus. To this end, the results clearly indicate that PmYRP65 is responsible for the receptor of the yellow head virus in P. monodon. Strikingly, the knockdown of the receptor is not lethal to the black tiger shrimp. The discovery strongly suggests that gene manipulation is applicable for the prevention of P. monodon from the contagion caused by the yellow head virus infection in shrimp farming industry.
About Wanchai Assavalapsakul
Wanchai Assalvalapsakul is an Assistant Professor in the Department of Microbiology at Chulalongkorn University in Thailand. His research interests include virology, molecular biology, Host-Virus interactions, the molecular biology of shrimp, and the cloning and expression of genes of interest. Wanchai is currently a visiting Research Fellow with QAAFI after being awarded an Endeavour Fellowship from the Australian Government.
Directions to UQ
- Google Map:
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- Directions:
- St Lucia Campus | Gatton campus.
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