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.

中文翻译：

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LTR衍生的miR-1269a和靶基因KSR2在schbasgelii中的表达分析。

背景技术Sebastes schlegelii是在韩国沿海地区发现的重要鱼类，具有重要的商业价值。迄今为止，大多数研究主要集中在环境因素上。行为模式，水产养殖，疾病和有限的基因研究，与microRNA（miRNA）或转座因子（TE）几乎没有关系。目的为了了解TE衍生的miR-1269a的生物学作用，我们检查了Sebastes schlegelii各个组织中miR-1269a及其靶基因KSR2的表达模式。另外，我们在HINAE细胞中进行了荧光素酶报告基因检测。方法使用UCSC Genome Browser（https://genome.ucsc.edu/）来检查哪些TE与miR-1269a相关。对于miR-1269a的靶基因，使用miRDB（http://www.mirdb.org/）和TargetScan 7.1（http://www.targetscan.org/vert_71/）鉴定与miRNA相关的靶基因。两步miRNA试剂盒，HB miR Multi Assay Kit™系统。我被用来分析TE衍生的miRNA表达模式。将KSR2基因的3'UTR克隆到psiCHECK-2载体中。随后与miR-1269a模拟物共转染至HINAE细胞，以进行萤光素酶报告基因检测。结果发现MiR-1269a来自LTR反转录转座子MLT2B。LTR衍生的miR-1269a在Sebastes schlegelii的肌肉，肝脏和性腺组织中高表达，而KSR2在脑中高表达。模仿HINAE细胞的KSR2和miR-1269a共转染显示出与KSR2相关的miR-1269a高活性。结论LTR衍生的miR-1269a在schbasgelii中显示出与KSR2相关的增强子活性。这些数据可作为进一步研究miRNA和靶基因相关性的基础，以及其他在施氏梭鲈中具有生物学意义的功能研究。