GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives

• DOI: 10.1016/j.stemcr.2016.07.015
• Publication Date: 2016-09-09T15:33:05Z
• Authors: Tim Kao; Tanya Labonne; Jonathan C. Niclis; Ritu Chaurasia; Zerina Lokmic; Elizabeth Qian; Freya F. Bruveris; Sara E. Howden; Ali Motazedian; Jacqueline V. Sch...; Magdaline Costa; Koula Sourris; Elizabeth Ng; David Anderson; Antonietta Giudice; Peter Farlie; Michael Cheung; Shireen R. Lamande; Anthony J. Penington; Clare L. Parish; Lachlan H. Thomson; Arash Rafii; David A. Elliott; Andrew G. Elefanty; Edouard G. Stanley
• Keywords: mCherry; Germ layer

The ability to reliably express fluorescent reporters or other genes of interest is important for using human pluripotent stem cells (hPSCs) as a platform investigating cell fates and gene function. We describe simple expression system, designated GAPTrap (GT), in which reporter genes, including GFP, mCherry, mTagBFP2, luc2, Gluc, lacZ are inserted into the GAPDH locus hPSCs. Independent clones harboring variations GT vectors expressed remarkably consistent levels gene. Differentiation experiments showed that was maintained hematopoietic cells, cardiac mesoderm, definitive endoderm, ventral midbrain dopaminergic neurons. Similarly, analysis teratomas derived from GT-lacZ hPSCs β-galactosidase spectrum types representing derivatives three germ layers. Thus, represent robust straightforward tagging system enables indelible labeling PSCs their differentiated derivatives.