Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Science (UIPS), Utrecht University, Utrecht, The Netherlands.
Methods Mol Biol. 2021;2355:275-286. doi: 10.1007/978-1-0716-1617-8_21.
Successful delivery of mRNA into the cytosol of professional antigen-presenting cells (APCs) poses one of the biggest challenges in developing effective mRNA vaccines to treat various cancers and viral infectious diseases. However, most polymeric mRNA delivery systems fail to transfect APCs. We have discovered that decoration of pH-sensitive endosome-disruptive GALA peptides on the surface of mRNA polyplexes leads to efficient targeting and transfection of APCs. GALA peptides not only enhance specific uptake in APCs through binding to sialic acid moieties, they also facilitate the endosomal escape of mRNA especially in dendritic cells (DCs). Here, we describe in detail the production of stabilized mRNA polyplexes post-conjugated with GALA peptides via copper-free click chemistry. Methods described here include the synthesis and purification of GALA peptides and its conjugation to mRNA polyplexes.
将 mRNA 递送到专业抗原呈递细胞 (APC) 的细胞质中是开发有效 mRNA 疫苗来治疗各种癌症和病毒感染性疾病的最大挑战之一。然而，大多数聚合物 mRNA 递送系统无法转染 APC。我们发现，在 mRNA 多聚物表面装饰 pH 敏感的内体破坏型 GALA 肽可有效靶向和转染 APC。GALA 肽不仅通过与唾液酸部分结合增强 APC 中的特异性摄取，还促进 mRNA 的内体逃逸，尤其是在树突状细胞 (DC) 中。在这里，我们通过无铜点击化学详细描述了通过 GALA 肽缀合后稳定化的 mRNA 多聚物的生产。这里描述的方法包括 GALA 肽的合成和纯化及其与 mRNA 多聚物的缀合。
