Gene delivery by a steroid-peptide nucleic acid conjugate.
We previously introduced a method called steroid-mediated gene delivery (SMGD), which uses steroid receptors as shuttles to facilitate the nuclear uptake of transfected DNA. Here, we describe a SMGD strategy with peptide nucleic acids (PNAs) that allowed linkage of a steroid molecule to a defined position in a plasmid without disturbing its gene expression. We synthesized and tested several bifunctional steroid derivatives [patent in process of nationalization] and finally selected the compound named DEX-bisPNA, a molecule consisting of a dexamethasone moiety linked to a PNA clamp (bisPNA) through a 30-atom chemical spacer. Dex-bisPNA binds to the glucocorticoid receptor (GR) as well as to reporter plasmids containing the corresponding PNA binding sites, translocates the GR from the cytoplasm into the nucleus, and increases the delivery of plasmid to the nucleus, resulting in enhanced GR-dependent expression of the reporter gene. The SMGD effect was more pronounced in growth-arrested cells than in proliferating cells. The specificity for the GR was shown by the reversion of the SMGD effect in the presence of dexamethasone as well as an enhanced expression in GR-positive cells but not in GR-negative cells. Thus, SMGD with PNA is a promising strategy for nonviral gene delivery into target tissues expressing specific steroid receptors.
Division of Nephrology and Hypertension and Clinical Research, University of Berne, CH-3010 Berne-Inselspital, Switzerland.
Alexandre G
AG
Rebuffat
Andrea R
AR
Nawrocki
Peter E
PE
Nielsen
Alessio G
AG
Bernasconi
Eloy
E
Bernal-Mendez
Brigitte M
BM
Frey
Felix J
FJ
Frey
D021581:Active Transport, Cell Nucleus; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D003907:Dexamethasone / Q000737:chemistry / Q000378:metabolism; D015195:Drug Design; D017930:Genes, Reporter; D005938:Glucocorticoids / Q000737:chemistry; D008954:Models, Biological; D009696:Nucleic Acids / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection / Q000379:methods*
D005938:Glucocorticoids; D009696:Nucleic Acids; D020135:Peptide Nucleic Acids; D011965:Receptors, Glucocorticoid; C464982:dexamethasone-bis-peptide nucleic acid; D003907:Dexamethasone