Details

Autor(en) / Beteiligte

• Patsch, Christoph
• Peitz, Michael
• Otte, David M.
• Kesseler, Daniela
• Jungverdorben, Johannes
• Wunderlich, Frank Thomas
• Brüstle, Oliver
• Zimmer, Andreas
• Edenhofer, Frank

Titel

Engineering Cell‐Permeant FLP Recombinase for Tightly Controlled Inducible and Reversible Overexpression in Embryonic Stem Cells

Ist Teil von

• Stem cells (Dayton, Ohio), 2010-05, Vol.28 (5), p.894-902

Ort / Verlag

Hoboken: Wiley Subscription Services, Inc., A Wiley Company

Erscheinungsjahr

2010

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Combined application of DNA recombinases Cre and FLP enables tightly controlled independent and/or sequential gene regulations. However, in practice, such dual recombinase strategies are hampered by the comparably low efficiency of the FLP recombinase. Here, we present the engineering of a recombinant cell‐permeant FLP protein (TAT‐FLP) that induces recombination in >75% of fibroblasts and mouse as well as human embryonic stem (ES) cells. We show that TAT‐FLP ideally complements the strength of cell‐permeant Cre recombinase for genetic engineering as exemplified by FLP‐ON‐Cre‐OFF, an inducible transgene expression cassette that enables tightly controlled expression in a reversible manner. We exemplify this concept by conditional overexpression of LacZ and the caudal‐related homeobox transcription factor CDX2. We expect our FLP transduction system to become widely useful for numerous genetic interventions addressing complex biological questions and the generation of transgene‐free therapeutically applicable ES cell‐derived cells. STEM CELLS 2010;28:894–902