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Video protocol

Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification

JoVE 3155 11/18/2011

Dec 2007 Nanog-GFP Transgenic Mouse


Nanog-GFP Transgenic Mouse
A tool for generation of germline-competent iPS cells

BRC No. 02290_STOCK Tg(Nanog-GFP,Puro)1Yam
Research Application : Stem Cell Research
Developmental Biology
Generation of iPS cells by using Nanog-GFP Tg mouse (Nature, 448, 313-317 (2007))


mn0712_0103 mn0712_0102
Germline-competent iPS cells from Nanog-GFP transgenic mouse.
Phase contrast (left) and fluorescence (right) micrographs are shown.
  Nanog gene is a good candidate marker for maintaining pluripotent differentiation, because of the loss of pluripotent epiblast in Nanog KO mice, the tendency of Nanog-null ES cells to differenciate spontaneously, and the capability of Nanog-overexpressed ES cells to maintain self-renewal independently of leukaemia inhibitory factor (LIF). Nanog-GFP transgene was generated by insertion of GFP-IRES-puromycin resistance gene (Puror) cassette into the 5′ untranslated region of a bacterial artificial chromosome (BAC) containing the mouse Nanog gene. ES cells with the Nanog-GFP transgene were positive for GFP, but became negative when differentiation was induced. Using this system, pluripotent stem cells competent for germline chimaeras can be induced from Nanog-GFP transgenic mouse fibroblasts by introduction of candidate genes for pluripotency.
The iPS cells will be available soon from RIKEN BRC Cell Bank.


Depositor : Dr. Shinya Yamanaka (Kyoto University)
Reference :  Nature (2007) 448:313-7.