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Conditional expression of Lodavin, an avidin-tagged LDL receptor, for biotin-mediated applications in vivo
Authors:Murugan Subramanian  Saarela Ulla  Airenne Kari  Shan Jingdong  Skovorodkin Ilya  Ylä-Herttuala Seppo  Vainio Seppo J
Affiliation:Oulu Centre for Cell-Matrix Research, Biocenter Oulu, Laboratory of Developmental Biology, University of Oulu, Oulu, Finland; Department of Medical Biochemistry and Molecular Biology, University of Oulu, Oulu, Finland.
Abstract:Lodavin represents an engineered fusion protein that consists of a cytoplasmic and a transmembrane domain of the human low‐density lipoprotein receptor coupled to an extracellular avidin monomer. Biotinylated compounds have been successfully targeted to Lodavin‐expressing cells that have been transduced by a Lodavin‐containing virus, and the targeting is based on the high affinity between biotin and avidin. We engineered a Rosa26 (R26R) knock‐in Lodavin mouse to develop biotin‐based applications such as targeted drug delivery, cell purification, and tissue imaging in vivo. A cDNA encoding Lodavin was inserted downstream of a floxed βgeo resistance gene in the R26R locus in embryonic stem cells, and a germ line‐derived R26RLodavin mouse line was generated. Efficient removal of the floxed βgeo cassette and conditional activation of Lodavin expression was achieved as a result of crossing the R26RLodavin mice with HoxB7‐Cre, Wnt4‐Cre, or Tie1‐Cre mice. In summary, the R26RLodavin mouse line may provide a useful tool for testing and developing applications with the aid of avidin and biotin interaction. genesis 50:693–699, 2012. © 2012 Wiley Periodicals, Inc.
Keywords:targeted drug delivery  cell purification  imaging  Rosa26 locus  kidney development
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