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Next-generation sequencing of testicular miRNAs in Atlantic cod

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Kjetil Aune


Genomics in Aquaculture 2011; Crete, 2011-09-14–2011-09-17

Lazado, Carlo C.; Nagasawa, Kazue; Fernandes, Jorge

Expression of the different genes is regulated not only at the transcriptional level but as well as by different mechanisms at the post-transcriptional level, including regulation by microRNAs (miRNAs). This large class of small non-coding RNAs, generally of around 21- 24 nucleotides in length exerts down-regulation ability, either by translation inhibition or by degradation mRNA, on target genes through complementary binding to their 3′-UTR regions. The importance of miRNAs in many physiological processes is now widely acknowledged. Current next-generation sequencing technologies enable in-depth characterization of the global repertoire of miRNAs. Though, many microRNAs have already been discovered and characterized, there is still a staggering number unaccounted, many of which with unknown function. In the case of teleosts, there is a paucity of information on the dynamics of miRNA in many physiological processes such as in development, growth and maturation. Hence, this study was conducted to identify and characterize the miRNAs in the testis of Atlantic cod by SOLiD next generation sequencing technology during its reproductive and non-reproductive stage in the maturation cycle. The small RNA from the total testicular RNA samples was purified by flashPAGE™ fractionation and converted to amplified libraries using the SOLiD™ Small RNA Expression Kit. The cDNA libraries containing the adaptor sequences were clonally amplified onto bead with emulsion PCR using SOLiD™ ePCR kit and were then sequenced on the SOLiD™ Analyzer. A differential miRNA transcriptional profile was observed between reproductive and non-reproductive stage of cod. Bioinformatic and experimental analyses are still ongoing to further identify and characterize miRNAs that are functionally important during sexual maturation of cod.