GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives
Tim Kao, Tanya Labonne, Jonathan C Niclis, Ritu Chaurasia, Zerina Lokmic, Elizabeth Qian, Freya F Bruveris, Sara E Howden, Ali Motazedian, Jacqueline V Schiesser, Magdaline Costa, Koula Sourris, Elizabeth Ng, David Anderson, Antonietta Giudice, Peter Farlie, Michael Cheung, Shireen R Lamande, Anthony J Penington, Clare L Parish Show all
STEM CELL REPORTS | CELL PRESS | Published : 2016
The ability to reliably express fluorescent reporters or other genes of interest is important for using human pluripotent stem cells (hPSCs) as a platform for investigating cell fates and gene function. We describe a simple expression system, designated GAPTrap (GT), in which reporter genes, including GFP, mCherry, mTagBFP2, luc2, Gluc, and lacZ are inserted into the GAPDH locus in hPSCs. Independent clones harboring variations of the GT vectors expressed remarkably consistent levels of the reporter gene. Differentiation experiments showed that reporter expression was reliably maintained in hematopoietic cells, cardiac mesoderm, definitive endoderm, and ventral midbrain dopaminergic neurons...View full abstract
Awarded by European Union
The authors thank Matt Burton and Paul Lau for help with flow cytometry and Katerina Vlahos for assistance with teratoma experiments. This work was supported by the Victorian Government's Operational Infrastructure Support Program and Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme (NHMRC IRIISS), Stem Cells Australia, the NHMRC, and the Victoria-California Stem Cell Alliance. We are also grateful for the generous support of the Stafford Fox Foundation. D.J.A. was supported by the European Union's Seventh Framework Program (FP7/2007-2013) under grant agreement PIOF-GA-2010-276186.). C.L.P. was supported by a Viertel Senior Medical Research Fellowship, Australia. A.G.E. and E.G.S. are Senior Research Fellows of the NHMRC. S.E.H. is supported by an NHMRC Overseas Biomedical Fellowship.