Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USA.
Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA.
Methods Mol Biol. 2024;2822:367-386. doi: 10.1007/978-1-0716-3918-4_23.
Transfection with mRNA has been considered superior to that with plasmids since the mRNA can be translated to a protein in the cytosol without entering the nucleus. One disadvantage of using mRNA is its susceptibility to enzymatic biodegradability, and consequently, significant research has occurred to determine nonviral carriers that will sufficiently stabilize this nucleic acid for cellular transport. Histidine-lysine peptides (HK) are one such class of mRNA carriers, which we think serves as a model for other peptides and polymeric carrier systems. When the HK peptide and mRNA are mixed and interact through ionic and nonionic bonds, mRNA polyplexes are formed, which can transfect cells. In contrast to linear HK peptides, branched HK peptides protected and efficiently transfected mRNA into cells. After describing the preparation and biophysical characterization of these polyplexes, we will provide protocols for in vitro and in vivo transfection for these mRNA polyplexes.
转染 mRNA 被认为优于转染质粒，因为 mRNA 可以在细胞质中翻译成蛋白质，而无需进入细胞核。使用 mRNA 的一个缺点是其易被酶降解，因此，人们进行了大量研究来确定非病毒载体，以充分稳定这种核酸，使其能够进行细胞运输。组氨酸-赖氨酸肽（HK）是一类 mRNA 载体，我们认为它可以作为其他肽和聚合载体系统的模型。当 HK 肽和 mRNA 混合并通过离子和非离子键相互作用时，就会形成 mRNA 多聚物，从而可以转染细胞。与线性 HK 肽相比，分支 HK 肽可以保护并有效地将 mRNA 转染到细胞中。在描述这些多聚物的制备和生物物理特性后，我们将提供这些 mRNA 多聚物的体外和体内转染方案。
