PvRaf knockdown using RNA interference increased survival of Litopenaeus vannamei infected with white spot syndrome virus
Joseph Carlo V. Vergel1, Mary Beth B. Maningas2.
Shrimp aquaculture is a major source of income in the intertropical countries of the Southeast Asian region. However, a great decline in production is observed due to a major viral pathogen, the White Spot Syndrome Virus (WSSV) which continues to prevail despite many preventive measures applied to deter the virus. RNA interference (RNAi) technology has been employed to reveal
functions of specific genes in the virus and its host with the aim of controlling WSSV by elucidating
complex host-virus interactions. This study determined the involvement of PvRaf, a key component of the Ras/Raf/MAPK pathway. Raf promotes cell survival through its protein-to-protein interactions and MEK-ERK independent mechanism. DNA sequencing revealed that PvRaf has a resulting size of 1.9 kb. Phylogenetic analysis revealed that PvRaf is highly homologous to the reported Raf genes of other organisms. Moreover, PvRaf is ubiquitously expressed in vital organs suggesting that it is essential to metabolic functions of the shrimp and may also play a role in its innate immune system as highlighted in its expression in the hemocytes. Quantitative Real-Time PCR (qRT-PCR) results confirmed the in vivo gene knockdown of PvRaf using RNA interference. qRT-PCR showed that PvRaf knockdown yields downregulation of the gene in the PvRaf-dsRNA and GFP-dsRNA treated shrimps. Gene expression analysis showed significant downregulation from day-0 to day-1 post-infection in PvRafdsRNA treatment relative to the control samples. Statistical analysis of the survival data indicated that PvRaf-dsRNA treatment has a significant protective effect against WSSV compared to GFP-dsRNA and PBS-treated shrimps.
Affiliation:
- National University Philippines, Philippines
- College of Science, University of Santo Tomas, Philippines, Philippines
Toggle translation
Download this article (This article has been downloaded 16 time(s))