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STTM 技术敲低 microRNA-143-5p 影响黑素细胞中真黑素和褐黑素的生成。

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

机构信息

College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi 030801, P.R. China.

出版信息

Mol Med Rep. 2019 Sep;20(3):2649-2656. doi: 10.3892/mmr.2019.10492. Epub 2019 Jul 12.

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.

摘要

微小 RNA(miRNAs)在哺乳动物黑素细胞的黑素生成调控中发挥多种作用,有助于头发颜色的形成。黑素细胞作用的调控是遗传改良的新目标。短串联靶标模拟物(STTM)是在植物和动物中沉默 miRNAs 的有效方法。为了研究 miR-143-5p 在黑素生成中的功能,使用 STTM 阻断 miR-143-5p 的表达(STTM-miR-143-5p)。分子分析和荧光素酶报告基因检测鉴定肌球蛋白 Va 基因(MYO5A)是 miR-143-5p 的靶标之一。STTM-miR-143-5p 的过表达导致下游黑素生成基因的表达增加,包括小眼相关转录因子(MITF)、酪氨酸酶家族成员[酪氨酸酶(TYR)和酪氨酸酶相关蛋白 1(TYRP1)]、黑素小体蛋白(MLPH)和 Rab27a,从而通过促进总碱溶性黑素生成(ASM)和真黑素(EM)含量促进黑素细胞色素沉着;相反,STTM-miR-143-5p 的过表达导致酪氨酸酶相关蛋白 2(TYRP2)/多巴色素互变异构酶(DCT)的表达降低,这导致小鼠黑素细胞中 pheomelanin(PM)含量降低。结果表明,通过使用 STTM 操纵 miR-143-5p 的表达,可以增加黑素细胞中的黑色素生成,从而导致 ASM 和 EM 的产生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1534/6691272/e4e0886432b7/MMR-20-03-2649-g00.jpg

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