Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Biosci Rep. 2018 Feb 21;38(1). doi: 10.1042/BSR20171160. Print 2018 Feb 28.
MicroRNAs (miRNAs) have important roles in many cellular processes, including cell proliferation, growth and development, and disease control. Previous study demonstrated that the expression of two highly homologous miRNAs (miR-192 and miR-215) was up-regulated in weaned piglets with F18 infection. However, the potential molecular mechanism of miR-192 in regulating infection remains unclear in pigs. In the present study, we analyzed the relationship between level of miR-192 and degree of resistance using transcription activator-like effector nuclease (TALEN), bacterial adhesion assays, and target genes research. A TALEN expression vector that specifically recognizes the pig miR-192 was constructed and then monoclonal epithelial cells defective in miR-192 were established. We found that miR-192 knockout led to enhance the adhesion ability of the strains F18ab, F18ac and K88ac, meanwhile increase the expression of target genes ( and ) by qPCR and Western blotting analysis. The results suggested that miR-192 and its key target genes ( and ) could have a key role in infection. Based on our findings, we propose that further investigation of miR-192 function is likely to lead to insights into the molecular mechanisms of infection.
微小 RNA（miRNAs）在许多细胞过程中发挥重要作用，包括细胞增殖、生长和发育以及疾病控制。先前的研究表明，两种高度同源的 miRNAs（miR-192 和 miR-215）在感染 F18 的断奶仔猪中表达上调。然而，miR-192 在调节猪感染中的潜在分子机制仍不清楚。在本研究中，我们使用转录激活样效应物核酸酶（TALEN）、细菌黏附试验和靶基因研究分析了 miR-192 水平与抗性程度之间的关系。构建了一种特异性识别猪 miR-192 的 TALEN 表达载体，然后建立了 miR-192 缺失的单克隆上皮细胞。我们发现 miR-192 敲除导致 F18ab、F18ac 和 K88ac 菌株的黏附能力增强，同时通过 qPCR 和 Western blot 分析增加了靶基因（和）的表达。结果表明，miR-192 及其关键靶基因（和）可能在 F18 感染中起关键作用。基于我们的发现，我们提出进一步研究 miR-192 的功能可能有助于深入了解 F18 感染的分子机制。
