Networking Centre on Bioengineering, Biomaterials and Nanomedicine (CIBER BBN), Barcelona, Spain.
Institute for Advanced Chemistry of Catalonia (IQAC), Spanish Research Council (CSIC), Barcelona, Spain.
Methods Mol Biol. 2021;2282:119-136. doi: 10.1007/978-1-0716-1298-9_8.
Nucleic acid conjugates are promising drugs for treating gene-related diseases. Conjugating specific units like lipids, cell-penetrating peptides, polymers, antibodies, and aptamers either at the 3'- or 5'-termini of a siRNA duplex molecule has resulted in a plethora of siRNA bioconjugates with improved stabilities in bloodstream and better pharmacokinetic values than unmodified siRNAs. In this sense, lipid-siRNA conjugates have attracted a remarkable interest for their potential value in facilitating cellular uptake. In this chapter, we describe a series of protocols involving the synthesis of siRNA oligonucleotides carrying either neutral or cationic lipids at the 3'- and 5'-termini. The resulting lipid-siRNA conjugates are aimed to be used as exogenous effectors for inhibiting gene expression by RNA interference. A protocol for the formulation of lipid siRNA using sonication in the presence of serum is described yielding interesting transfection properties for cell culture without the use of transfecting agents.
核酸偶联物是治疗基因相关疾病的有前途的药物。在 siRNA 双链体分子的 3'或 5'末端连接特定的单元，如脂质、细胞穿透肽、聚合物、抗体和适体，已经产生了大量的 siRNA 生物缀合物，其在血液中的稳定性得到改善，药代动力学值优于未修饰的 siRNA。从这个意义上说，脂质-siRNA 缀合物因其在促进细胞摄取方面的潜在价值而引起了极大的关注。在本章中，我们描述了一系列涉及合成带有中性或阳离子脂质的 siRNA 寡核苷酸的方案。所得的脂质-siRNA 缀合物旨在用作通过 RNA 干扰抑制基因表达的外源性效应物。描述了一种使用血清存在下的超声处理来制备脂质 siRNA 的方案，该方案在不使用转染剂的情况下对细胞培养具有有趣的转染特性。
