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Expression analysis of LTR-derived miR-1269a and target gene, KSR2 in Sebastes schlegelii

  • Jennifer Im
  • Woo Ryung Kim
  • Hee-Eun Lee
  • Ahran Kim
  • Do-Hyung Kim
  • Yung Hyun Choi
  • Hee-Jae Cha
  • Suhkmann Kim
  • Heui-Soo KimEmail author
Research Article
  • 82 Downloads

Abstract

Background

Sebastes schlegelii are an important species of fish found in the coastal areas of the Korea with significant commercial importance. Most studies thus far have been primarily focused on environmental factors; behavioural patterns, aquaculture, diseases and limited genetic studies with little to none related to either microRNAs (miRNAs) or transposable elements (TE).

Objectives

In order to understand biological roles of TE-derived miR-1269a, we examined expression pattern for miR-1269a and its target gene, KSR2, in various tissues of Sebastes schlegelii. Also, we performed luciferase reporter assay in HINAE cells.

Methods

UCSC Genome Browser (https://genome.ucsc.edu/) was used to examine which TE is associated with miR-1269a. For the target genes for miR-1269a, the target genes associated with the miRNA were identified using miRDB (http://www.mirdb.org/) and TargetScan 7.1 (http://www.targetscan.org/vert_71/). A two-step miRNA kit, HB miR Multi Assay Kit™ System. I was used for the analysis of TE-derived miRNA expression patterns. The 3′UTR of KSR2 gene was cloned into the psiCHECK-2 vector. Subsequently co-transfected with miR-1269a mimics to HINAE cells for luciferase reporter assay.

Results

MiR-1269a was found to be derived from LTR retrotransposon, MLT2B. LTR-derived miR-1269a was highly expressed in the muscle, liver and gonad tissues of Sebastes schlegelii, but KSR2 revealed high expression in the brain. Co-transfection of KSR2 and miR-1269a mimic to HINAE cells showed high activity of miR-1269a in relation to KSR2.

Conclusion

LTR-derived miR-1269a showed enhancer activity with relation to KSR2 in Sebastes schlegelii. The data may be used as a foundation for further investigation regarding correlation of miRNA and target genes in addition to other functional studies of biological significance in Sebastes schlegelii.

Keywords

miR-1269a KSR2 Transposable element Sebastes schlegelii MLT2B 

Notes

Acknowledgements

This research was a part of the project titled “Omics based on fishery disease control technology development and industrialization (20150242) funded by the Ministry of Oceans and Fisheries, Korea.

Funding

This research was a part of the project titled “Omics based on fishery disease control technology development and industrialization (20150242) funded by the Ministry of Oceans and Fisheries, Korea.

Compliance with ethical standards

Conflict of interest

Jennifer Im, Woo Ryung Kim, Hee-Eun Lee, Ahran Kim, Do-Hyung Kim, Yung Hyun Choi, Hee-Jae Cha, Suhkmann Kim and Heui-Soo Kim declare that we have no conflict of interest.

Ethical approval

All experiments were carried out in accordance with the guidelines and regulation approved by Pusan National University-Institutional Animal Care and Use Committee (PNU-IACUC).

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Copyright information

© The Genetics Society of Korea 2019

Authors and Affiliations

  1. 1.Department of Biological Sciences, College of Natural SciencesPusan National UniversityBusanRepublic of Korea
  2. 2.Institute of Systems BiologyPusan National UniversityBusanRepublic of Korea
  3. 3.Department of Aquatic Life Medicine, College of Fisheries SciencesPukyong National UniversityBusanRepublic of Korea
  4. 4.Department of Biochemistry, College of Korean MedicineDong-eui UniversityBusanRepublic of Korea
  5. 5.Department of Parasitology and Genetics, College of MedicineKosin UniversityBusanRepublic of Korea
  6. 6.Department of Chemistry, College of Natural SciencesPusan National UniversityBusanRepublic of Korea

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