Characterization of Argonaute cDNA from Penaeus monodon and implication of its role in RNA interference.
RNA interference (RNAi) has recently become a promising strategy for therapeutic of several viral diseases including those in the black tiger shrimp Penaeus monodon. However, the protein components that play role in RNAi in P. monodon have not yet been identified. Here, we report the cloning and functional characterization of a cDNA encoding Argonaute, a principal constituent of RNAi pathway in P. monodon. P. monodon's Argonaute (Pem-AGO) exhibited the two signature domains, PAZ and PIWI. Substantial level of Pem-ago expression could be suppressed by double-stranded RNA (dsRNA) that targeted PAZ coding sequence in shrimp primary culture of Oka cells. The Pem-ago depleted cells showed impaired RNAi as the expression of an endogenous gene was rescued from the dsRNA-mediated silencing in these cells. Our results imply that Pem-ago is required for effective RNAi in P. monodon and thus identify the first protein constituent of RNAi machinery in penaeid shrimp.
Dechklar
Manasave
M
Institute of Molecular Biology and Genetics, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya Campus, Nakhon Pathom 73170, Thailand.
Udomkit
Apinunt
A
Panyim
Sakol
S
D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics*; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D034622:RNA Interference
D018076:DNA, Complementary; D011506:Proteins