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http://dx.doi.org/10.5657/FAS.2014.0377

Simultaneous and Systemic Knock-down of Big Defensin 1 and 2 gene Expression in the Pacific Oyster Crassostrea gigas using Long Double-stranded RNA-mediated RNA Interference  

Jee, Bo Young (Aquatic Life Disease Control Division, National Fisheries Research and Development Institute)
Kim, Min Sun (Department of Aquatic Life Medicine, Pukyong National University)
Cho, Mi Young (Aquatic Life Disease Control Division, National Fisheries Research and Development Institute)
Lee, Soon Jeong (Aquatic Life Disease Control Division, National Fisheries Research and Development Institute)
Park, Myung Ae (Aquatic Life Disease Control Division, National Fisheries Research and Development Institute)
Kim, Jin Woo (Pathology division, National Fisheries Research and Development Institute)
Choi, Seung Hyuk (Department of Aquatic Life Medicine, Pukyong National University)
Jeong, Hyun Do (Department of Aquatic Life Medicine, Pukyong National University)
Kim, Ki Hong (Department of Aquatic Life Medicine, Pukyong National University)
Publication Information
Fisheries and Aquatic Sciences / v.17, no.3, 2014 , pp. 377-380 More about this Journal
Abstract
RNA interference (RNAi)-mediated transcriptional knock-down of Crassostrea gigas big defensin 1 and 2 genes (Cg-BigDef1 and Cg-BigDef2) was investigated. The cDNA sequences of Cg-BigDef1 and Cg-BigDef2 were identical, excluding an additional fragment of 20 nucleotides in Cg-BigDef1; thus, a long double-stranded RNA (dsRNA) targeting the mRNA of Cg-BigDef2 effectively downregulated both Cg-BigDef2 and Cg-BigDef1. In addition, long dsRNA targeting green fluorescent protein (GFP) did not affect transcription of the two big defensin genes. These results suggest that the transcriptional downregulation of Cg-BigDef1 and Cg-BigDef2 was mediated by sequence-specific RNA interference (RNAi). Despite injection of long dsRNA targeting Cg-BigDef2 into only the adductor muscle, knock-down of Cg-BigDef1 and Cg-BigDef2 was observed in the adductor muscle, hemocytes, mantle, and gills, suggestive of systemic spread of RNAi in C. gigas. Furthermore, the inhibitory effect of dsRNA persisted until 72 h post-injection, indicative of a long-lasting RNAi-mediated knock-down of target genes.
Keywords
Crassostrea gigas; Long double-stranded RNA; RNA interference; Big defensins;
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