Department of Pharmacology, Ehime University Graduate School of Medicine, Toon, Ehime, Japan.
Methods Mol Biol. 2024;2766:145-151. doi: 10.1007/978-1-0716-3682-4_15.
RNA interference (RNAi) is a widely used technique to regulate the expression of genes and proteins with a high degree of specificity that is not easily accessed by traditional pharmacological approaches. For preclinical research on rheumatoid arthritis (RA), silencing of target genes in primary immune cells can be easily achieved by the application of small interfering RNA (siRNA) and synthetic short hairpin RNA (shRNA). Cellular and systemic administration of siRNA or shRNA has been a significant advance in preclinical research on RA. In this chapter, the basic techniques for gene silencing in human-derived peripheral T cells using liposome-dependent siRNA transfection and lentiviral-mediated shRNA delivery, aiming at gene silencing of therapeutic targets, are introduced.
RNA 干扰 (RNAi) 是一种广泛用于调节基因和蛋白质表达的技术,具有高度特异性,这是传统药理学方法难以实现的。对于类风湿关节炎 (RA) 的临床前研究,通过应用小干扰 RNA (siRNA) 和合成短发夹 RNA (shRNA),可以轻松实现靶基因在原代免疫细胞中的沉默。siRNA 或 shRNA 的细胞内和全身给药在 RA 的临床前研究中是一项重大进展。在本章中,介绍了使用脂质体依赖性 siRNA 转染和慢病毒介导的 shRNA 递送在人源外周 T 细胞中进行基因沉默的基本技术,目的是实现治疗靶点的基因沉默。