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. TSI通风表/多参数通风表
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