Sawai K, Ohno K, Iijima Y, Levin B, Meruelo D
Department of Pathology, New York University Medical Center, New York 10016, USA.
Mol Genet Metab. 1998 May;64(1):44-51. doi: 10.1006/mgme.1998.2692.
In this study, we developed a cell-specific mRNA transfection system using streptavidin-protein A (ST-PA) fusion protein and monoclonal antibodies (mAbs). We previously reported that ST-PA fusion protein and mAb complexes can transfer certain biotinylated proteins into specific cell types. At this time, we combined an in vitro transcribed biotinylated and self-replicating Sindbis virus genomic RNA with ST-PA fusion protein and mAbs. In the presence of cationic liposomes, to prevent RNA degradation, this complex is able to transfect a reporter gene to specific cancer cells in a mAb does-dependent manner. Even in the absence of cationic liposomes, biotinylated mRNA, ST-PA fusion, and mAb complexes can transfer some types of cancer cell suspension cultures. This cell-specific transfection system is a novel method of introducing various mRNAs into cells that results in high levels of transient protein expression.
在本研究中，我们利用链霉亲和素-蛋白A（ST-PA）融合蛋白和单克隆抗体（mAb）开发了一种细胞特异性mRNA转染系统。我们之前报道过，ST-PA融合蛋白和mAb复合物可以将某些生物素化蛋白转移到特定细胞类型中。此时，我们将体外转录的生物素化且自我复制的辛德毕斯病毒基因组RNA与ST-PA融合蛋白和mAb相结合。在阳离子脂质体存在的情况下，为防止RNA降解，该复合物能够以mAb剂量依赖性方式将报告基因转染至特定癌细胞。即使在没有阳离子脂质体的情况下，生物素化mRNA、ST-PA融合物和mAb复合物也可以转染某些类型的癌细胞悬浮培养物。这种细胞特异性转染系统是一种将各种mRNA导入细胞的新方法，可导致高水平的瞬时蛋白表达。
