CAS Key Laboratory of Pathogenic Microbiology and Immunology (CASPMI), Institute of Microbiology, Chinese Academy of Sciences, Beijing, People's Republic of China.
PLoS One. 2011;6(11):e26380. doi: 10.1371/journal.pone.0026380. Epub 2011 Nov 11.
The transfer of foreign genes into mammalian cells has been essential for understanding the functions of genes and mechanisms of genetic diseases, for the production of coding proteins and for gene therapy applications. Currently, the identification and selection of cells that have received transferred genetic material can be accomplished by methods, including drug selection, reporter enzyme detection and GFP imaging. These methods may confer antibiotic resistance, or be disruptive, or require special equipment. In this study, we labeled genetically modified cells with a cell surface biotinylation tag by co-transfecting cells with BirA, a biotin ligase. The modified cells can be quickly isolated for downstream applications using a simple streptavidin bead method. This system can also be used to screen cells expressing two sets of genes from separate vectors.
将外源基因转入哺乳动物细胞对于理解基因的功能和遗传疾病的机制、编码蛋白的生产以及基因治疗的应用至关重要。目前，可以通过包括药物选择、报告酶检测和 GFP 成像在内的方法来鉴定和选择已接收转移遗传物质的细胞。这些方法可能会赋予抗生素抗性，或者具有破坏性，或者需要特殊设备。在这项研究中，我们通过共转染 BirA（一种生物素连接酶）将细胞表面生物素化标签与基因修饰细胞进行标记。通过使用简单的链霉亲和素珠方法，可以快速分离修饰后的细胞，用于下游应用。该系统还可用于筛选来自单独载体的两组基因表达的细胞。
