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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.

Generation of a constitutively expressing Tetracycline repressor (TetR) human embryonic stem cell line BJNhem20-TetR