Selective enhancement of gene transfer by steroid-mediated gene delivery.
The incorporation of transgenes into the host cells' nuclei is problematic using conventional nonviral gene delivery technologies. Here we describe a strategy called steroid-mediated gene delivery (SMGD), which uses steroid receptors as shuttles to facilitate the uptake of transfected DNA into the nucleus. We use glucocorticoid receptors (GRs) as a model system with which to test the principle of SMGD. To this end, we synthesized and tested several bifunctional steroid derivatives, finally focusing on a compound named DR9NP, consisting of a dexamethasone backbone linked to a psoralen moiety using a nine-atom chemical spacer. DR9NP binds to the GR in either its free or DNA-crosslinked form, inducing the translocation of the GR to the nucleus. The expression of transfected DR9NP-decorated reporter plasmids is enhanced in dividing cells: expression of steroid-decorated reporter plasmids depends on the presence of the GR, is independent of the transactivation potential of the GR, and correlates with enhanced nuclear accumulation of the transgene in GR-positive cells. The SMGD effect is also observed in cells naturally expressing GRs and is significantly increased in nondividing cell cultures. We propose that SMGD could be used as a platform for selective targeting of transgenes in nonviral somatic gene transfer.
Rebuffat
A
A
Division of Nephrology, Inselspital, Bern, Switzerland.
Bernasconi
A
A
Ceppi
M
M
Wehrli
H
H
Verca
S B
SB
Ibrahim
M
M
Frey
B M
BM
Frey
F J
FJ
Rusconi
S
S
D021581:Active Transport, Cell Nucleus; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005363:Ficusin / Q000737:chemistry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008024:Ligands; D018613:Microscopy, Confocal; D008954:Models, Biological; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D013256:Steroids / Q000378:metabolism*; D014162:Transfection; D019076:Transgenes; D001616:beta-Galactosidase / Q000378:metabolism
D003432:Cross-Linking Reagents; D008024:Ligands; D011965:Receptors, Glucocorticoid; D013256:Steroids; D004247:DNA; D001616:beta-Galactosidase; D005363:Ficusin