Abstract
Protein transduction domains (PTDs), such as the HIV1-TAT peptide, have been previously used to promote the uptake of proteins into a range of cell types, including stem cells. Here we generated pancreatic transcription factors containing PTD sequences and administered these to endoderm enriched mouse embryonic stem (ES) cells under conditions that were designed to mimic the pattern of expression of these factors in the developing pancreas. The ES cells were first cultured as embryoid bodies and treated with Activin A and Bone morphogenetic protein 4 (BMP4) to promote formation of definitive endoderm. Cells were subsequently plated as a monolayer and treated with different combinations of the modified recombinant transcription factors Pdx1 and MafA. The results demonstrate that each transcription factor was efficiently taken up by the cells, where they were localized in the nuclei. RT-qPCR was used to measure the expression levels of pancreatic markers. After the addition of Pdx1 alone for a period of five days, followed by the combination of Pdx1 and TAT-MafA in a second phase, up-regulation of insulin 1, insulin 2, Pdx1, Glut2, Pax4 and Nkx6.1 was observed. As assessed by immunocytochemistry, double positive insulin and Pdx1 ...Continue Reading
References
Dec 23, 1988·Cell·A D Frankel, C O Pabo
Oct 13, 1994·Nature·J JonssonH Edlund
Jan 18, 1994·Proceedings of the National Academy of Sciences of the United States of America·S FawellJ Barsoum
Feb 27, 1999·Experimental Cell Research·M V Wiles, B M Johansson
Jul 27, 2000·The New England Journal of Medicine·A M ShapiroR V Rajotte
May 9, 2001·Nucleic Acids Research·M W Pfaffl
Nov 3, 2001·Diabetologia·C M McKinnon, K Docherty
Jun 28, 2003·Diabetes·Hirofumi NoguchiSusan Bonner-Weir
Mar 5, 2004·Development·Atsushi KuboGordon Keller
Mar 1, 2005·Diabetes·Juan Domínguez-BendalaHelena Edlund
Jun 29, 2005·Diabetes·Edmond A RyanA M James Shapiro
Aug 30, 2005·Developmental Biology·Michael A HaleRaymond J MacDonald
Nov 1, 2005·Nature Biotechnology·Kevin A D'AmourEmmanuel E Baetge
Apr 4, 2006·Developmental Biology·Wataru NishimuraArun Sharma
Jul 28, 2006·Biochemical and Biophysical Research Communications·Jing ChenDaru Lu
Mar 6, 2007·Developmental Cell·Kerstin A JohanssonAnne Grapin-Botton
Mar 24, 2007·Analytical Biochemistry·Aziza ManceurJulie Audet
Oct 5, 2007·Trends in Molecular Medicine·Jacob M Gump, Steven F Dowdy
Oct 26, 2007·Seminars in Cell & Developmental Biology·Kevin DochertyLudovic Vallier
Dec 19, 2007·Diabetes·Vijay KoyaLi-Jun Yang
Feb 22, 2008·Nature Biotechnology·Evert KroonEmmanuel E Baetge
Oct 18, 2008·Diabetes, Obesity & Metabolism·E E Baetge
Nov 18, 2008·Developmental Biology·George K Gittes
Dec 6, 2008·Stem Cells·Andreia S BernardoKevin Docherty
Jul 16, 2010·Diabetes·Isabella ArtnerRoland Stein
Sep 30, 2010·Cell Transplantation·M SundbergE Sundström
Citations
Jun 8, 2018·PloS One·Yanan DongYukun Li
Dec 3, 2013·Stem Cells Translational Medicine·Taku KaitsukaKazuhito Tomizawa
Jul 14, 2016·Stem Cells Translational Medicine·Elisa CorritorePhilippe A Lysy
Jun 22, 2017·Frontiers in Genetics·Andhira VieiraPatrick Collombat
Jan 27, 2019·Stem Cells·Ioannis GianniosAnthony Gavalas
Dec 15, 2020·Protein Expression and Purification·Gloria NarayanRajkumar P Thummer