Department of Dermatology, Indiana University School of Medicine, 545 Barnhill Dr. Emerson 139, Indianapolis, IN, 46202, USA.
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN, USA.
Sci Rep. 2020 Sep 23;10(1):15517. doi: 10.1038/s41598-020-72027-1.
Human papillomavirus (HPV) L1 and L2 capsid proteins self-assemble into virions capable of efficiently packaging either its 8 kilobase genome or non-viral DNA. The ability of HPV capsids to package non-viral DNA makes these a useful tool for delivering plasmids to study proteins of interest in a variety of cell types. We describe optimization of current methods and present new protocols for using HPV capsids to deliver non-viral DNA thereby providing an alternative to DNA transfection. Using keratinocyte generated extracellular matrices can enhance infection efficiency in keratinocytes, hepatocytes and neuronal cells. Furthermore, we describe a suspension-based efficient technique for infecting different cell types.
人乳头瘤病毒 (HPV) L1 和 L2 衣壳蛋白自行组装成病毒粒子,能够有效地包装其 8 千碱基对基因组或非病毒 DNA。HPV 衣壳包装非病毒 DNA 的能力使这些病毒成为将质粒递送到各种细胞类型中研究感兴趣蛋白质的有用工具。我们描述了当前方法的优化,并提出了使用 HPV 衣壳递非病毒 DNA 的新方案,从而为 DNA 转染提供了替代方法。使用角质形成细胞生成的细胞外基质可以提高角质形成细胞、肝细胞和神经元细胞的感染效率。此外,我们还描述了一种基于悬浮液的高效感染不同细胞类型的技术。