title: Real-time in vivo imaging of stem cells following transgenesis by transposition.
abstract: Previous studies have identified Sleeping Beauty transposons as efficient vectors for nonviral gene delivery in mammalian cells. However, studies demonstrating the usefulness of transposons as gene delivery vehicles into adult stem cells are lacking. Multipotent adult progenitor cells (MAPC) are nonhematopoietic stem cells with the capacity to form most, if not all, cell types of the body and as such hold great therapeutic potential. The whole-body biodistribution and persistence of MAPC are unknown, and such data would help direct clinical applications. We have nucleofected murine MAPC with two plasmid-based Sleeping Beauty transposons encoding the red fluorescent protein (DsRed2) and firefly luciferase. Transgenic euploid MAPC clones maintained their characteristic multilineage differentiation potential in vitro. DsRed2 and luciferase expression allowed for MAPC detection in vivo and in tissue sections. To confirm that transgenesis occurred by transposition into the genome of MAPC, we mapped Sleeping Beauty transposon integration sites in two MAPC clones using splinkerette PCR. This novel dual-reporter imaging approach based on the transgenesis of MAPC with Sleeping Beauty transposons sheds light on the homing patterns of MAPC and paves the way for quantification of MAPC engraftment in real time in vivo.
authors.affiliation: Department of Pediatrics, Division of Hematology, Oncology, Blood and Marrow Transplant Program, University of Minnesota, Minneapolis, MN 55455, USA. tolar003@umn.edu
authors.forename: Jakub
authors.initials: J
authors.lastname: Tolar
authors.forename: Mark
authors.initials: M
authors.lastname: Osborn
authors.forename: Scott
authors.initials: S
authors.lastname: Bell
authors.forename: Ron
authors.initials: R
authors.lastname: McElmurry
authors.forename: Lily
authors.initials: L
authors.lastname: Xia
authors.forename: Megan
authors.initials: M
authors.lastname: Riddle
authors.forename: Angela
authors.initials: A
authors.lastname: Panoskaltsis-Mortari
authors.forename: Yuehua
authors.initials: Y
authors.lastname: Jiang
authors.forename: R Scott
authors.initials: RS
authors.lastname: McIvor
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Contag
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Yant
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kay
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Verfaillie
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Blazar
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements; D003952:Diagnostic Imaging; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D008156:Luciferases; D008164:Luminescent Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D039902:Multipotent Stem Cells
chemical_list: D004251:DNA Transposable Elements; D008164:Luminescent Proteins; C413663:fluorescent protein 583; D008156:Luciferases