Combination of let-7d-5p, miR-26a-5p, and miR-15a-5p is suitable normalizer for studying microRNA expression in skin tissue of Liaoning cashmere goat during hair follicle cycle W.L., Dang Y.L., Yin R.H., Yin R.L., Jiang W.Q., Wang Z.Y., Zhu Y.B., Wang J.J., Zhao Z.H., Luo G.B. (2016): Combination of let-7d-5p, miR-26a-5p, and miR-15a-5p is suitable normalizer for studying microRNA expression in skin tissue of Liaoning cashmere goat during hair follicle cycle. Czech J. Anim. Sci., 61: 99-107.
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The microRNAs are non-coding RNA molecules of approximately 20–22 nucleotides that are found to be implicated in a wide range of physiological processes. In this study, the suitability of 10 candidate reference RNAs was evaluated for microRNA expression data in the skin tissue of Liaoning cashmere goat including 1 small nuclear RNA (snRNA; RNU6B), 1 small nucleolar RNA (snoRNA; Z30), 1 rRNA (5S), 1 transfer RNA (tRNA; Met-tRNA), and 6 microRNAs (miR; let-7d-5p, miR-15a-5p, miR-26a-5p, miR-125a-5p, miR-214-3p, and miR-221-3p). Based on geNorm and NormFinder algorithms, we identified let-7d-5p, miR-26a-5p, and miR-15a-5p as the most stable reference RNAs. Also, three reference RNAs (let-7d-5p, miR-26a-5p, and miR-15a-5p) were sufficient for the normalization of microRNA expression data in the skin of this breed. We further assessed the suitability of let-7d-5p, miR-26a-5p, and miR-15a-5p in a combination as reference RNAs through detecting the relative expression of miR-24-3p, miR-29a-3p, miR-145a-5p, and miR-205-5p as putative genes of interest. Significant differences were revealed in the relative expression of miR-24-3p, miR-29a-3p, miR-145a-5p, and miR-205-5p at telogen stage of hair follicle cycle when a combination of let-7d-5p, miR-26a-5p, and miR-15a-5p vs a single let-7d-5p were used as reference RNA. Based on the results from this study, we suggested that the combination of let-7d-5p, miR-26a-5p, and miR-15a-5p as normalizers for microRNA expression data would be more reliable than that of single let-7d-5p, and the geometric mean of these three microRNAs (let-7d-5p, miR-26a-5p, and miR-15a-5p) can be used for the normalization of microRNAs expression data in the skin of Liaoning cashmere goat.
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