Cite this paper:
PU Longjun, WANG Jing, WANG Yu, ZUO Jianwei, GUO Huarong. Computational identification and characterization of microRNAs and their targets in Penaeus monodon[J]. Journal of Oceanology and Limnology, 2018, 36(3): 853-869

Computational identification and characterization of microRNAs and their targets in Penaeus monodon

PU Longjun1, WANG Jing1, WANG Yu1, ZUO Jianwei1, GUO Huarong1,2
1 Ministry of Education Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China;
2 Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, China
Abstract:
Study on shrimp miRNAs was limited and just 7 mature miRNA sequences of Marsupenaeus japonicus are deposited in mirBase database. In this study, miRNAs and their target gene candidates were computationally identified from shrimp Penaeus monodon and then experimentally validated. Using 39 908 expressed sequence tags (ESTs) and 21 124 genome survey sequences (GSSs) of P. monodon (pmo) as reference dataset, a comprehensive approach based on inter-species homolog search was employed to investigate the candidate miRNAs (i.e. pmo-miRNA). A total of eight miRNAs belonging to 7 families were computationally identified and five out of them were subsequently validated by PCR and sequencing. Of these, pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-3819 were first identified from shrimps. Both the mature pmo-miRNAs and the corresponding precursors were conserved among different species. Based on perfect or near-perfect match to the target region, the target gene candidates of pmomiRNAs were predicted from 10 331 mRNA sequences of P. monodon. A total of 20 genes were predicted as the targets of pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-6492. Experimental validation by dual luciferase reporter assay confirmed the targeting between 3 pmo-miRNAs and one or two of their target genes, especially the pmo-miR-4979 which could significantly down-regulate the expression of target gene (JR226772). This study updates the miRNAs and their targets in P. monodon and lays a solid foundation for future RNAi study.
Key words:    microRNA|shrimp|Penaeus monodon|target genes   
Received: 2016-12-26   Revised:
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