Publication

DNA nuclear targeting sequences for non-viral gene delivery

van Gaal, E. V. B., Oosting, R. S., van Eijk, R., Bakowska, M., Feyen, D., Kok, R. J., Hennink, W. E., Crommelin, D. J. A. & Mastrobattista, E., Jul-2011, In : Pharmaceutical Research. 28, 7, p. 1707-22 16 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

van Gaal, E. V. B., Oosting, R. S., van Eijk, R., Bakowska, M., Feyen, D., Kok, R. J., ... Mastrobattista, E. (2011). DNA nuclear targeting sequences for non-viral gene delivery. Pharmaceutical Research, 28(7), 1707-22. https://doi.org/10.1007/s11095-011-0407-8

Author

van Gaal, Ethlinn V B ; Oosting, Ronald S ; van Eijk, Roel ; Bakowska, Marta ; Feyen, Dries ; Kok, Robbert Jan ; Hennink, Wim E ; Crommelin, Daan J A ; Mastrobattista, Enrico. / DNA nuclear targeting sequences for non-viral gene delivery. In: Pharmaceutical Research. 2011 ; Vol. 28, No. 7. pp. 1707-22.

Harvard

van Gaal, EVB, Oosting, RS, van Eijk, R, Bakowska, M, Feyen, D, Kok, RJ, Hennink, WE, Crommelin, DJA & Mastrobattista, E 2011, 'DNA nuclear targeting sequences for non-viral gene delivery', Pharmaceutical Research, vol. 28, no. 7, pp. 1707-22. https://doi.org/10.1007/s11095-011-0407-8

Standard

DNA nuclear targeting sequences for non-viral gene delivery. / van Gaal, Ethlinn V B; Oosting, Ronald S; van Eijk, Roel; Bakowska, Marta; Feyen, Dries; Kok, Robbert Jan; Hennink, Wim E; Crommelin, Daan J A; Mastrobattista, Enrico.

In: Pharmaceutical Research, Vol. 28, No. 7, 07.2011, p. 1707-22.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

van Gaal EVB, Oosting RS, van Eijk R, Bakowska M, Feyen D, Kok RJ et al. DNA nuclear targeting sequences for non-viral gene delivery. Pharmaceutical Research. 2011 Jul;28(7):1707-22. https://doi.org/10.1007/s11095-011-0407-8


BibTeX

@article{a02cbd5e77b541fbb57d30449682c281,
title = "DNA nuclear targeting sequences for non-viral gene delivery",
abstract = "PURPOSE: To evaluate if introduction of DNA nuclear Targeting Sequences (DTS; i.e. recognition sequences for endogenous DNA-binding proteins) in plasmid DNA (pDNA) leads to increased transfection efficiency of non-viral gene delivery by virtue of enhanced nuclear import of the pDNA.METHODS: A set of DTS was identified and cloned into EGFP-reporter plasmids controlled by the CMV-promoter. These pDNA constructs were delivered into A431 and HeLa cells using standard electroporation, pEI-based polyfection or lipofection methods. The amount of pDNA delivered into the nucleus was determined by qPCR; transfection efficiency was determined by flow cytometry.RESULTS: Neither of these DTS increased transgene expression. We varied several parameters (mitotic activity, applied dose and delivery strategy), but without effect. Although upregulated transgene expression was observed after stimulation with TNF-α, this effect could be ascribed to non-specific upregulation of transcription rather than enhanced nuclear import. Nuclear copy numbers of plasmids containing or lacking a DTS did not differ significantly after lipofectamine-based transfection in dividing and non-dividing cells.CONCLUSION: No beneficial effects of DTS on gene expression or nuclear uptake were observed in this study.",
keywords = "Base Sequence, Cell Line, Tumor, Cell Nucleus, Electroporation, Flow Cytometry, Gene Transfer Techniques, Genetic Vectors, HeLa Cells, Humans, Molecular Sequence Data, Plasmids, Polymerase Chain Reaction",
author = "{van Gaal}, {Ethlinn V B} and Oosting, {Ronald S} and {van Eijk}, Roel and Marta Bakowska and Dries Feyen and Kok, {Robbert Jan} and Hennink, {Wim E} and Crommelin, {Daan J A} and Enrico Mastrobattista",
year = "2011",
month = "7",
doi = "10.1007/s11095-011-0407-8",
language = "English",
volume = "28",
pages = "1707--22",
journal = "Pharmaceutical Research",
issn = "0724-8741",
publisher = "SPRINGER/PLENUM PUBLISHERS",
number = "7",

}

RIS

TY - JOUR

T1 - DNA nuclear targeting sequences for non-viral gene delivery

AU - van Gaal, Ethlinn V B

AU - Oosting, Ronald S

AU - van Eijk, Roel

AU - Bakowska, Marta

AU - Feyen, Dries

AU - Kok, Robbert Jan

AU - Hennink, Wim E

AU - Crommelin, Daan J A

AU - Mastrobattista, Enrico

PY - 2011/7

Y1 - 2011/7

N2 - PURPOSE: To evaluate if introduction of DNA nuclear Targeting Sequences (DTS; i.e. recognition sequences for endogenous DNA-binding proteins) in plasmid DNA (pDNA) leads to increased transfection efficiency of non-viral gene delivery by virtue of enhanced nuclear import of the pDNA.METHODS: A set of DTS was identified and cloned into EGFP-reporter plasmids controlled by the CMV-promoter. These pDNA constructs were delivered into A431 and HeLa cells using standard electroporation, pEI-based polyfection or lipofection methods. The amount of pDNA delivered into the nucleus was determined by qPCR; transfection efficiency was determined by flow cytometry.RESULTS: Neither of these DTS increased transgene expression. We varied several parameters (mitotic activity, applied dose and delivery strategy), but without effect. Although upregulated transgene expression was observed after stimulation with TNF-α, this effect could be ascribed to non-specific upregulation of transcription rather than enhanced nuclear import. Nuclear copy numbers of plasmids containing or lacking a DTS did not differ significantly after lipofectamine-based transfection in dividing and non-dividing cells.CONCLUSION: No beneficial effects of DTS on gene expression or nuclear uptake were observed in this study.

AB - PURPOSE: To evaluate if introduction of DNA nuclear Targeting Sequences (DTS; i.e. recognition sequences for endogenous DNA-binding proteins) in plasmid DNA (pDNA) leads to increased transfection efficiency of non-viral gene delivery by virtue of enhanced nuclear import of the pDNA.METHODS: A set of DTS was identified and cloned into EGFP-reporter plasmids controlled by the CMV-promoter. These pDNA constructs were delivered into A431 and HeLa cells using standard electroporation, pEI-based polyfection or lipofection methods. The amount of pDNA delivered into the nucleus was determined by qPCR; transfection efficiency was determined by flow cytometry.RESULTS: Neither of these DTS increased transgene expression. We varied several parameters (mitotic activity, applied dose and delivery strategy), but without effect. Although upregulated transgene expression was observed after stimulation with TNF-α, this effect could be ascribed to non-specific upregulation of transcription rather than enhanced nuclear import. Nuclear copy numbers of plasmids containing or lacking a DTS did not differ significantly after lipofectamine-based transfection in dividing and non-dividing cells.CONCLUSION: No beneficial effects of DTS on gene expression or nuclear uptake were observed in this study.

KW - Base Sequence

KW - Cell Line, Tumor

KW - Cell Nucleus

KW - Electroporation

KW - Flow Cytometry

KW - Gene Transfer Techniques

KW - Genetic Vectors

KW - HeLa Cells

KW - Humans

KW - Molecular Sequence Data

KW - Plasmids

KW - Polymerase Chain Reaction

U2 - 10.1007/s11095-011-0407-8

DO - 10.1007/s11095-011-0407-8

M3 - Article

VL - 28

SP - 1707

EP - 1722

JO - Pharmaceutical Research

JF - Pharmaceutical Research

SN - 0724-8741

IS - 7

ER -

ID: 19628927