title: Light-controlled modulation of gene expression using polyamidoamine formulations.
abstract: A promising method that offers both time- and site-specific delivery of macromolecules is photochemical internalization technology (PCI). Here, we have characterized various polyamidoamine (PAMAM) carriers [generation (G) 0-7], for light-directed delivery of nucleic acids in vitro by the use of PCI technology. A number of parameters for optimal delivery of nucleic acids into human cancer cells, that is, various light-doses, carrier-doses, and small interfering RNA (siRNA)/messenger RNA (mRNA) doses were investigated for either up- or down-regulation of enhanced green fluorescent protein (EGFP) gene expression. In summary, our results showed in an osteosarcoma cell line (OHS) [EGFP] model system the possibility for efficient light-directed siRNA silencing (>80% silencing) when using PAMAM G3 to G7 as carriers. Surprisingly, no EGFP mRNA up-regulation was detected either with or without PCI after EGFP mRNA/PAMAM (G0-G7) transfection in standard OHS cells. We have here identified properties for PAMAM formulations enabling light-directed siRNA delivery with the aim of developing a site-specific strategy for delivery of nucleic acids in vivo.
authors.affiliation: Department of Tumor Biology, Institute for Cancer Research , The Norwegian Radium Hospital, Montebello, Norway. sigurdbo@rr-research.no
authors.forename: Sigurd Leinæs
authors.initials: SL
authors.lastname: Bøe
authors.forename: Jens Andreas Lindin
authors.initials: JA
authors.lastname: Jørgensen
authors.forename: Ane Sager
authors.initials: AS
authors.lastname: Longva
authors.forename: Timothy
authors.initials: T
authors.lastname: Lavelle
authors.forename: Stein
authors.initials: S
authors.lastname: Sæbøe-Larssen
authors.forename: Eivind
authors.initials: E
authors.lastname: Hovig
mesh_terms: D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins; D006801:Humans; D008027:Light; D012516:Osteosarcoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D055668:Photochemical Processes; D017319:Photosensitizing Agents / Q000008:administration & dosage / Q000737:chemistry; D011073:Polyamines / Q000008:administration & dosage* / Q000737:chemistry; D012333:RNA, Messenger / Q000008:administration & dosage* / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry; D015854:Up-Regulation
chemical_list: D017319:Photosensitizing Agents; C531249:Poly(amidoamine); D011073:Polyamines; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins