Coupling of a targeting peptide to plasmid DNA by covalent triple helix formation

Carole Neves; Gerardo Byk; Daniel Scherman; Pierre Wils

doi:10.1016/s0014-5793(99)00674-2

Coupling of a targeting peptide to plasmid DNA by covalent triple helix formation

Carole Neves, Gerardo Byk, Daniel Scherman, Pierre Wils

Sanofi-Aventis

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

73 Scopus citations

Abstract

The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear entry of proteins. We have developed a strategy for covalent coupling of one or a controlled number of NLS peptides to plasmid DNA at a specific site by triple helix formation. A psoralen-oligonucleotide-NLS peptide conjugate was synthesized and characterized by proteolysis with trypsin. This conjugate was used to covalently associate one NLS peptide to plasmid DNA by triple helix formation and photoactivation. The oligonucleotide-NLS peptide conjugate interacted with the NLS-receptor importin α. The reporter gene was expressed after transfection of the modified plasmid in NIH 3T3 cells, indicating no loss of the gene expression functionality of the plasmid. On the other hand, no increase in expression was observed as a result of the NLS peptide. This site-specific coupling technology can be used to couple to a plasmid other ligands targeting to a specific receptor.

Original language	English
Pages (from-to)	41-45
Number of pages	5
Journal	FEBS Letters
Volume	453
Issue number	1-2
DOIs	https://doi.org/10.1016/s0014-5793(99)00674-2
State	Published - 18 Jun 1999
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by a grant from the French Ministry of Research. We thank Dr. François F. Clerc and his staff for peptide synthesis and Dr. Marc Vuilhorgne and his staff from the Structural Analysis Department of Rhône-Poulenc Rorer for mass spectroscopy analysis. We thank Dr. Y. Yoneda for the gift of the pGEX-2T plasmid. We thank D. Bisch, Dr. F. Blanche and Dr. V. Thuillier for importin α-GST purification.

Funding

This work was supported by a grant from the French Ministry of Research. We thank Dr. François F. Clerc and his staff for peptide synthesis and Dr. Marc Vuilhorgne and his staff from the Structural Analysis Department of Rhône-Poulenc Rorer for mass spectroscopy analysis. We thank Dr. Y. Yoneda for the gift of the pGEX-2T plasmid. We thank D. Bisch, Dr. F. Blanche and Dr. V. Thuillier for importin α-GST purification.

Funders	Funder number
French Ministry of Research

Keywords

DNA
Gene therapy
Gene transfer
Importin
Nuclear localization signal
Triple helix

Access to Document

10.1016/s0014-5793(99)00674-2

Cite this

@article{eb32d82d94ab4ebb8d9a4305a1c04073,

title = "Coupling of a targeting peptide to plasmid DNA by covalent triple helix formation",

abstract = "The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear entry of proteins. We have developed a strategy for covalent coupling of one or a controlled number of NLS peptides to plasmid DNA at a specific site by triple helix formation. A psoralen-oligonucleotide-NLS peptide conjugate was synthesized and characterized by proteolysis with trypsin. This conjugate was used to covalently associate one NLS peptide to plasmid DNA by triple helix formation and photoactivation. The oligonucleotide-NLS peptide conjugate interacted with the NLS-receptor importin α. The reporter gene was expressed after transfection of the modified plasmid in NIH 3T3 cells, indicating no loss of the gene expression functionality of the plasmid. On the other hand, no increase in expression was observed as a result of the NLS peptide. This site-specific coupling technology can be used to couple to a plasmid other ligands targeting to a specific receptor.",

keywords = "DNA, Gene therapy, Gene transfer, Importin, Nuclear localization signal, Triple helix",

author = "Carole Neves and Gerardo Byk and Daniel Scherman and Pierre Wils",

note = "Funding Information: This work was supported by a grant from the French Ministry of Research. We thank Dr. Fran{\c c}ois F. Clerc and his staff for peptide synthesis and Dr. Marc Vuilhorgne and his staff from the Structural Analysis Department of Rh{\^o}ne-Poulenc Rorer for mass spectroscopy analysis. We thank Dr. Y. Yoneda for the gift of the pGEX-2T plasmid. We thank D. Bisch, Dr. F. Blanche and Dr. V. Thuillier for importin α-GST purification.",

year = "1999",

month = jun,

day = "18",

doi = "10.1016/s0014-5793(99)00674-2",

language = "אנגלית",

volume = "453",

pages = "41--45",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "John Wiley and Sons Inc.",

number = "1-2",

}

TY - JOUR

T1 - Coupling of a targeting peptide to plasmid DNA by covalent triple helix formation

AU - Neves, Carole

AU - Byk, Gerardo

AU - Scherman, Daniel

AU - Wils, Pierre

N1 - Funding Information: This work was supported by a grant from the French Ministry of Research. We thank Dr. François F. Clerc and his staff for peptide synthesis and Dr. Marc Vuilhorgne and his staff from the Structural Analysis Department of Rhône-Poulenc Rorer for mass spectroscopy analysis. We thank Dr. Y. Yoneda for the gift of the pGEX-2T plasmid. We thank D. Bisch, Dr. F. Blanche and Dr. V. Thuillier for importin α-GST purification.

PY - 1999/6/18

Y1 - 1999/6/18

N2 - The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear entry of proteins. We have developed a strategy for covalent coupling of one or a controlled number of NLS peptides to plasmid DNA at a specific site by triple helix formation. A psoralen-oligonucleotide-NLS peptide conjugate was synthesized and characterized by proteolysis with trypsin. This conjugate was used to covalently associate one NLS peptide to plasmid DNA by triple helix formation and photoactivation. The oligonucleotide-NLS peptide conjugate interacted with the NLS-receptor importin α. The reporter gene was expressed after transfection of the modified plasmid in NIH 3T3 cells, indicating no loss of the gene expression functionality of the plasmid. On the other hand, no increase in expression was observed as a result of the NLS peptide. This site-specific coupling technology can be used to couple to a plasmid other ligands targeting to a specific receptor.

AB - The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear entry of proteins. We have developed a strategy for covalent coupling of one or a controlled number of NLS peptides to plasmid DNA at a specific site by triple helix formation. A psoralen-oligonucleotide-NLS peptide conjugate was synthesized and characterized by proteolysis with trypsin. This conjugate was used to covalently associate one NLS peptide to plasmid DNA by triple helix formation and photoactivation. The oligonucleotide-NLS peptide conjugate interacted with the NLS-receptor importin α. The reporter gene was expressed after transfection of the modified plasmid in NIH 3T3 cells, indicating no loss of the gene expression functionality of the plasmid. On the other hand, no increase in expression was observed as a result of the NLS peptide. This site-specific coupling technology can be used to couple to a plasmid other ligands targeting to a specific receptor.

KW - DNA

KW - Gene therapy

KW - Gene transfer

KW - Importin

KW - Nuclear localization signal

KW - Triple helix

UR - http://www.scopus.com/inward/record.url?scp=0032806151&partnerID=8YFLogxK

U2 - 10.1016/s0014-5793(99)00674-2

DO - 10.1016/s0014-5793(99)00674-2

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

C2 - 10403371

AN - SCOPUS:0032806151

SN - 0014-5793

VL - 453

SP - 41

EP - 45

JO - FEBS Letters

JF - FEBS Letters

IS - 1-2

ER -

Coupling of a targeting peptide to plasmid DNA by covalent triple helix formation

Abstract

Bibliographical note

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Cite this