Knockdown of microRNA-143-5p by STTM technology affects eumelanin and pheomelanin production in melanocytes

Abstract

MicroRNAs (miRNAs) serve various roles in the regulation of melanogenesis in mammalian melanocytes that contribute to the development of hair color. The manipulation of the melanocyte action is a new target for genetic improvement. Short tandem target mimic (STTM) is a potent approach for silencing miRNAs in plants and animals. To investigate the function of miR-143-5p in melanogenesis, STTM was used to block the expression of miR-143-5p (STTM-miR-143-5p). The molecular analysis and luciferase reporter assay identified myosin Va gene (MYO5A) as one of the miR-143-5p targets. STTM-miR-143-5p overexpression resulted in an increased expression of downstream melanogenesis genes including microphthalmia-associated transcription factor (MITF), tyrosinase family members [tyrosinase (TYR) and tyrosinase-related protein 1 (TYRP1)], melanophilin (MLPH), and Rab27a, thereby contributing to melanocyte pigmentation by promoting total alkali-soluble melanogenesis (ASM) and eumelanin (EM) contents; conversely, STTM-miR-143-5p overexpression resulted in decreased expression of the tyrosinase-related protein 2 (TYRP2)/dopachrome tautomerase (DCT), which is responsible for decreased pheomelanin (PM) content in mouse melanocytes. The results indicated that melanin production in melanocytes could be increased by manipulating miR-143-5p expression using STTM which resulted in ASM and EM production.