Shetty Ronak, Inamdar Maneesha S
Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India.
Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India; Institute for Stem Cell Biology and Regenerative Medicine, Bengaluru, India.
Stem Cell Res. 2016 Mar;16(2):271-3. doi: 10.1016/j.scr.2015.12.043. Epub 2016 Jan 6.
Human embryonic stem cell line BJNhem20-TetR was generated using non-viral method. The construct pCAG-TetRnls was transfected using microporation procedure. BJNhem20-TetR can subsequently be transfected with any vector harbouring a TetO (Tet operator) sequence to generate doxycycline based inducible line. For example, in human embryonic stem cells, the pSuperior based TetO system has been transfected into a TetR containing line to generate OCT4 knockdown cell line (Zafarana et al., 2009). Thus BJNhem20-TetR can be used as a tool to perturb gene expression in human embryonic stem cells.
人类胚胎干细胞系BJNhem20-TetR是采用非病毒方法构建的。使用微孔转染程序转染构建体pCAG-TetRnls。随后，BJNhem20-TetR可以用任何含有TetO（四环素操纵子）序列的载体进行转染，以生成基于强力霉素的诱导细胞系。例如，在人类胚胎干细胞中，基于pSuperior的TetO系统已被转染到一个含有TetR的细胞系中，以生成OCT4基因敲低细胞系（扎法拉纳等人，2009年）。因此，BJNhem20-TetR可作为一种工具来干扰人类胚胎干细胞中的基因表达。
