Discovery of two novel miRNAs from the Ovis aries by a combinatorial approach of experiments and bioinformatics

Abstract

In the study, we successful constructed a library from the ovine testis using next-generation sequencing technology, and identified 219 novel miRNAs by bioinformatics. Two of the novel miRNAs (ovis_aries_testis-m0052_5p a nd ovis_aries_testis-m0165_5p), which were expressed in the sheep testis and ovary, and were confirmed by real-time PCR and northern blotting. Ovis_aries_testis-m0052_5p was 23 nucleotides in length, was located on chromosome 15, and had 100% similarity to mmu-miR-34c-5p, hsa-miR-34c-5p, gga-miR-34c-5p, and cfa-miR-34c. Ovis_aries_testis-m0165_5p was 21 nucleotides in length, located on chromosome 5, and had 100% similarity to mmu-miR-145a-5p, hsa-miR-145-5p, ssc-miR-145-5p, and bta-miR-145. The pre-miRNAs for Ovis_aries_testis-m0052_5p and Ovis_aries_testis-m0165_5p were 75 and 81 nucleotides in length, and both had a standard hairpin stem-loop structure. From the consistency of the sequence and structure, we speculated that ovis_aries_testis-m0052_5p had a function similar to hsa-miR-34c-5p, mmumiR-34c-5p, and ovis_aries_testis-m0165_5p had a function similar to hsa-miR-145-5p, which were involved in the fine regulation of cell survival, spermatozoon generate, breeding activities. Therefore, we defined the identified miRNAs as oar-miR-34c-5p and oar-miR-145-5p. The results will enrich the miRNA database, and provide the basis for research into the regulatory mechanisms of miRNA in relation to breeding activities.

Comparative rna‐seq analysis of region‐specific miRNA expression in the epididymis of cattleyak

“…These findings illustrate that miR‐145 might play some important roles in the male infertility, and sperm maturation or sperm motility of bovine epididymis. Another miRNA, namely miR‐145‐5p, has been shown in the regulation of oocyte maturation and could be clinically employed to improve the developmental potential of immature oocytes from infertile couples undergoing IVM (Chang et al., 2018). Recent study has suggested that miR‐ 145‐5p may play a pivotal role in inducing embryonic development, sex regulation, sperm production, fine regulation of cell survival, spermatozoon production and breeding activities (Cui et al., 2018).…”

​Re-sequencing of Whole Genome Jordanian Awassi Ewes using Hiseq Sequencing Technology: Major Step Towards Sheep Genomic Selection

“…Alberto et al (2018) determined certain genomic regions carrying genes involved in the nervous system, immunity and productivity traits differentiating domestic breeds of sheep and goats from wild populations. Chang et al (2018), reported two novel miRNAs from the Ovis aries may play an important role in inducing embryonic development, sex regulation, production of sperm and ovulation. Moreover, Chang et al (2020), identified few genes serve as potential biomarkers for monitoring disease progression and abortion risk during pregnancy in sheep.…”

Identification of Circular RNAs in the Ovaries of Hu Sheep and Local Fat-tailed Sheep during the Luteal Phase

“…In recent years, with the development of high-throughput sequencing, novel bioinformatics methods and related experimental validation, several classes of non-coding RNAs, such as circRNA, lncRNA and miRNA, have emerged and appear to play an important role in various biological processes (Chang et al, 2018;He et al, 2019;Chen et al, 2020;Zhou et al, 2020;Mei et al, 2021;Wu et al, 2021). CircRNA is different from linear RNA without 5 cap and 3 end poly-A tail.…”