Conjugation of a New Two-Photon Fluorophore to Poly(ethylenimine) for Gene Delivery Imaging
| Autor: | Patrice L. Baldeck, Ali Hayek, Sebnem Ercelen, Xin Zhang, Emmanuel Schaub, Frédéric Bolze, Jean-François Nicoud, Yves Mely |
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| Přispěvatelé: | Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Masson, Beatrice, Institut de Physique et Chimie des Matériaux de Strasbourg (IPCMS), Barthel, Ingrid |
| Rok vydání: | 2007 |
| Předmět: |
Fluorophore
Biomedical Engineering [SDV.BBM.BP] Life Sciences [q-bio]/Biochemistry Molecular Biology/Biophysics Pharmaceutical Science Bioengineering Fluorescence correlation spectroscopy macromolecular substances Gene delivery Transfection chemistry.chemical_compound Isothiocyanates Humans Polyethyleneimine Organic chemistry Fluorescent Dyes Pharmacology Fluorenes Photons Phenyl Ethers Spectrum Analysis Organic Chemistry Gene Transfer Techniques technology industry and agriculture DNA Chromophore Fluorescence Endocytosis [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biophysics Microscopy Fluorescence chemistry Biophysics HeLa Cells Biotechnology Conjugate |
| Zdroj: | Bioconj. Chem. Bioconj. Chem., 2007, 18, pp.844-851 Bioconjugate Chem. Bioconjugate Chem., 2007, 18, pp.844-851 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/bc060362h |
| Popis: | We report herein the molecular engineering of an efficient two-photon absorbing (TPA) chromophore based on a donor-donor bis-stilbenyl entity to allow conjugation with biologically relevant molecules. The dye has been functionalized using an isothiocyanate moiety to conjugate it with the amine functions of poly(ethylenimine) (PEI), which is a cationic polymer commonly used for nonviral gene delivery. Upon conjugation, the basic architecture and photophysical properties of the active TPA chromophore remain unchanged. At the usual N/P ratio (ratio of the PEI positive charges to the DNA negative charges) of 10 used for transfection, the transfection efficiency and cytotoxicity of the labeled PEI/DNA complexes were found to be comparable to those of the unlabeled PEI/DNA complexes. Moreover, when used in combination with unlabeled PEI (at a ratio of 1 labeled PEI to 3 unlabeled PEI), the labeled PEI does not affect the size of the complexes with DNA. The labeled PEI was successfully used in two-photon fluorescence correlation spectroscopy measurements, showing that at N/P = 10 most PEI molecules are free and the diffusion coefficient of the complexes is consistent with the 360 nm size measured by quasielastic light scattering. Finally, two-photon images of the labeled PEI/DNA complexes confirmed that the complexes enter into the cytoplasm of HeLa cells by endocytosis and hardly escape from the endosomes. As a consequence, the functionalized TPA chromophore appears to be an adequate tool to label the numerous polyamines used in nonviral gene delivery and characterize their complexes with DNA in two-photon applications. |
| Databáze: | OpenAIRE |
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