一种用于转染的 RNA 稳定化方法，可控制表达持续时间。
A method for stabilizing RNA for transfection that allows control of expression duration.
机构信息
Department of Biological Structure, Institute for Stem Cells and Regenerative Medicine, University of Washington School of Medicine, 815 Mercer Street, Seattle, WA 98109, USA.
出版信息
Dev Dyn. 2010 Jul;239(7):2034-40. doi: 10.1002/dvdy.22344.
RNA transfection methods have not proven to be as popular as DNA methods due to the highly transient nature of the RNA inside the cell. However, there are many advantages in using RNA for gene over-expression, such as the rapidity of expression, the ability to express in all cell types without the need for cell-type-specific promoters, and the ability to analyze the effects of gene over-expression in a transient manner. Therefore, we have developed a method (StabiLizingUtr: SLU) to stabilize the RNA for varying durations, using specific sequences from the 3'UTR of the Venezuelan equine encephalitis virus (VEEV). We have designed a plasmid for cloning genes upstream from repeated stabilizing sequences to generate mRNA with one or more VEEV-stabilizing sequence motifs. We demonstrate this method in several cell and tissue types, including the mammalian cochlea, a tissue that has been difficult to transfect with other methods.
RNA 转染方法由于细胞内 RNA 的高度瞬时性质，尚未像 DNA 方法那样受欢迎。然而，使用 RNA 进行基因过表达有许多优势，例如表达的快速性，无需细胞类型特异性启动子即可在所有细胞类型中表达的能力，以及以瞬时方式分析基因过表达的影响的能力。因此，我们开发了一种方法（StabiLizingUtr：SLU），使用委内瑞拉马脑炎病毒（VEEV）3'UTR 的特定序列来稳定 RNA 的不同持续时间。我们设计了一种质粒，用于在重复稳定序列的上游克隆基因，以产生具有一个或多个 VEEV 稳定序列基序的 mRNA。我们在几种细胞和组织类型中证明了这种方法，包括哺乳动物耳蜗，这是一种用其他方法难以转染的组织。
Zotero 插件