Purdue Institute for Integrative Neuroscience, Purdue University, West Lafayette, IN 47906, USA.
Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
BMB Rep. 2019 May;52(5):324-329. doi: 10.5483/BMBRep.2019.52.5.195.
Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols [BMB Reports 2019; 52(5): 324-329].
细胞重编程技术和谱系特异性细胞分化的最新进展为转化研究提供了巨大的机会。由于基于病毒的基因传递不是一种实用的重编程方案，基于蛋白质的重编程作为一种产生重编程细胞的安全方法受到了关注。然而，重编程蛋白的细胞摄取效率仍然是一个主要障碍。在这里，我们报道了可提高这些蛋白细胞摄取的关键因素。与 9 个赖氨酸（dsRED-9K）融合的纯化红色荧光蛋白作为细胞穿透肽可有效递送至多种原代细胞。加入氨酚喹可提高蛋白传递效率。此外，纯化的 dsRED-9K 能够有效地穿透源自小鼠胚胎干细胞的所有细胞谱系。我们的数据可能为基于蛋白质的重编程或分化方案的设计提供重要的见解。[BMB 报告 2019；52(5): 324-329]。
