Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex

Lionel Marcélis; Nancy Van Overstraeten; Javiera Lambermont; Sébastien Bontems; Nicolas Spinelli; Eric Defrancq; Cécile Moucheron; Andrée Kirsch De Mesmaeker; Martine Raes

doi:10.1002/cplu.201402212

Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex

Lionel Marcélis, Nancy Van Overstraeten, Javiera Lambermont, Sébastien Bontems, Nicolas Spinelli, Eric Defrancq, Cécile Moucheron, Andrée Kirsch De Mesmaeker, Martine Raes

Laboratory of Cellular Biochemistry and Biology

Research output: Contribution to journal › Article › peer-review

Abstract

A photoreactive ruthenium(II) complex that contains two tetraazaphenanthrene (TAP) and one phenanthroline (phen) ligands was synthesized and then tethered to (antisense) oligonucleotides (Ru-ASO) to target a destabilized GFP (dGFP). The specificity of the photoreaction of this Ru-ASO conjugate was studied in vitro by polyacrylamide gel electrophoresis (PAGE) experiments in denaturing conditions. Other nonspecific Ru-ASO conjugates were also prepared and evaluated with human keratinocytes that expressed dGFP. An illumination-dependent cytotoxicity was observed for most Ru-ASO conjugates that varied from 10 to almost 40%, but only the specific Ru-ASO conjugate was able to significantly reduce GFP expression in illuminated cells.

Original language	English
Pages (from-to)	1597-1604
Number of pages	8
Journal	ChemPlusChem
Volume	79
Issue number	11
DOIs	https://doi.org/10.1002/cplu.201402212
Publication status	Published - 2014

Keywords

Antisense agents
Electron transfer
Oligonucleotides
Photochemistry
Ruthenium

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10.1002/cplu.201402212

Cite this

Marcélis, L., Van Overstraeten, N., Lambermont, J., Bontems, S., Spinelli, N., Defrancq, E., Moucheron, C., Mesmaeker, A. K. D., & Raes, M. (2014). Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex. ChemPlusChem, 79(11), 1597-1604. https://doi.org/10.1002/cplu.201402212

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title = "Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex",

abstract = "A photoreactive ruthenium(II) complex that contains two tetraazaphenanthrene (TAP) and one phenanthroline (phen) ligands was synthesized and then tethered to (antisense) oligonucleotides (Ru-ASO) to target a destabilized GFP (dGFP). The specificity of the photoreaction of this Ru-ASO conjugate was studied in vitro by polyacrylamide gel electrophoresis (PAGE) experiments in denaturing conditions. Other nonspecific Ru-ASO conjugates were also prepared and evaluated with human keratinocytes that expressed dGFP. An illumination-dependent cytotoxicity was observed for most Ru-ASO conjugates that varied from 10 to almost 40%, but only the specific Ru-ASO conjugate was able to significantly reduce GFP expression in illuminated cells.",

keywords = "Antisense agents, Electron transfer, Oligonucleotides, Photochemistry, Ruthenium",

author = "Lionel Marc{\'e}lis and {Van Overstraeten}, Nancy and Javiera Lambermont and S{\'e}bastien Bontems and Nicolas Spinelli and Eric Defrancq and C{\'e}cile Moucheron and Mesmaeker, {Andr{\'e}e Kirsch De} and Martine Raes",

year = "2014",

doi = "10.1002/cplu.201402212",

language = "English",

volume = "79",

pages = "1597--1604",

journal = "ChemPlusChem",

issn = "2192-6506",

publisher = "Wiley-VCH Verlag",

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Marcélis, L, Van Overstraeten, N, Lambermont, J, Bontems, S, Spinelli, N, Defrancq, E, Moucheron, C, Mesmaeker, AKD & Raes, M 2014, 'Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex', ChemPlusChem, vol. 79, no. 11, pp. 1597-1604. https://doi.org/10.1002/cplu.201402212

TY - JOUR

T1 - Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex

AU - Marcélis, Lionel

AU - Van Overstraeten, Nancy

AU - Lambermont, Javiera

AU - Bontems, Sébastien

AU - Spinelli, Nicolas

AU - Defrancq, Eric

AU - Moucheron, Cécile

AU - Mesmaeker, Andrée Kirsch De

AU - Raes, Martine

PY - 2014

Y1 - 2014

N2 - A photoreactive ruthenium(II) complex that contains two tetraazaphenanthrene (TAP) and one phenanthroline (phen) ligands was synthesized and then tethered to (antisense) oligonucleotides (Ru-ASO) to target a destabilized GFP (dGFP). The specificity of the photoreaction of this Ru-ASO conjugate was studied in vitro by polyacrylamide gel electrophoresis (PAGE) experiments in denaturing conditions. Other nonspecific Ru-ASO conjugates were also prepared and evaluated with human keratinocytes that expressed dGFP. An illumination-dependent cytotoxicity was observed for most Ru-ASO conjugates that varied from 10 to almost 40%, but only the specific Ru-ASO conjugate was able to significantly reduce GFP expression in illuminated cells.

AB - A photoreactive ruthenium(II) complex that contains two tetraazaphenanthrene (TAP) and one phenanthroline (phen) ligands was synthesized and then tethered to (antisense) oligonucleotides (Ru-ASO) to target a destabilized GFP (dGFP). The specificity of the photoreaction of this Ru-ASO conjugate was studied in vitro by polyacrylamide gel electrophoresis (PAGE) experiments in denaturing conditions. Other nonspecific Ru-ASO conjugates were also prepared and evaluated with human keratinocytes that expressed dGFP. An illumination-dependent cytotoxicity was observed for most Ru-ASO conjugates that varied from 10 to almost 40%, but only the specific Ru-ASO conjugate was able to significantly reduce GFP expression in illuminated cells.

KW - Antisense agents

KW - Electron transfer

KW - Oligonucleotides

KW - Photochemistry

KW - Ruthenium

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U2 - 10.1002/cplu.201402212

DO - 10.1002/cplu.201402212

M3 - Article

AN - SCOPUS:84919429947

SN - 2192-6506

VL - 79

SP - 1597

EP - 1604

JO - ChemPlusChem

JF - ChemPlusChem

IS - 11

ER -

Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex

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Light-triggered green fluorescent protein silencing in human keratinocytes in culture using antisense oligonucleotides coupled to a photoreactive ruthenium(II) complex

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