Sequence-Specific Photomodification of Single-Stranded and Double-Stranded DNA Fragments by Oligonucleotide Perfluoroarylazide Derivative

Левина, А. С.; Tabatadze, David; Dobrikov, Mikhail I.; Shishkin, G. V.; Zarytova, Valentina P.

doi:10.1089/oli.1.1996.6.127

Cited by 11 publications

(7 citation statements)

References 18 publications

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“…Several methods for covalent attachment of molecules to nucleic acids have been developed and reported so far [11,12,15,16]. Among them, the use of photoactive molecules leads to efficient but non‐specific covalent modification of DNA [12,17]. A suitable method for site‐specific labeling of plasmid DNA involves the formation of a triple helix consisting of an oligonucleotide bound within the major groove of a double‐stranded target sequence [18].…”

Section: Discussionmentioning

confidence: 99%

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

et al. 1999

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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 K K. 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.z 1999 Federation of European Biochemical Societies.

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Section: Discussionmentioning

confidence: 99%

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

et al. 1999

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“…Several methods for covalent attachment of molecules to nucleic acids have been developed and reported so far (18)(19)(20). Among them, the use of photoactive molecules, such as tetrafluorophenylazido groups, leads to an efficient photomodification of DNA (21). We have conjugated a cationic peptide, NLS from SV40 large T antigen, to a photoactive tetrafluorophenylazido-containing molecule.…”

Section: Discussionmentioning

confidence: 99%

Coupling of Nuclear Localization Signals to Plasmid DNA and Specific Interaction of the Conjugates with Importin α

et al. 1998

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The nuclear localization signal (NLS) of the SV40 large T antigen efficiently induces nuclear targeting of proteins. We have developed a chemical strategy for covalent coupling of NLS peptides to plasmid DNA. A p-azido-tetrafluoro-benzyl-NLS peptide conjugate was synthesized. This conjugate was used to covalently associate NLS peptides to plasmid DNA by photoactivation. Reporter gene was expressed after transfection of the plasmid-NLS conjugates in NIH 3T3 cells. The conjugates interacted specifically with the NLS-receptor importin alpha, but plasmid-NLS conjugates were not detected in the nucleus, by fluorescence microscopy, after cytoplasmic microinjection.

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“…Preparation of Plasmid DNA − NLS Peptide Conjugates. Diazocoupling ( − ) and photoactiviation ( − ) have been reported as the covalent modifications of DNA. In this study plasmid DNA−NLS peptide conjugates were prepared by the diazo coupling method according with Scheme .…”

Section: Experimental Methodsmentioning

confidence: 99%

Can Nuclear Localization Signals Enhance Nuclear Localization of Plasmid DNA?

et al. 2003

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Nonviral vectors are safer and more cost-effective than viral vectors but are significantly less efficient, and thus, increasing the efficiency of nonviral vectors remains an important objective. One way to overcome this problem is by stimulating the nuclear localization of exogenous genes. Nuclear localization signals (NLSs) are known to be involved in the active transport of exogenous proteins and probes into the nucleus. However, stimulation of nuclear localization of plasmid DNA has yet to be confirmed completely. In the present study, we prepared plasmid DNA-NLS peptide conjugates and adjusted spacer length and number introduced in an attempt to increase transfection efficiency. In comparison to conjugates with unmodified plasmid DNA and short spacers, we found that NLS-plasmid DNA conjugates with covalent bonding by diazo coupling through PEG chain (MW 3400) stimulated complexation with the nuclear transport proteins importin alpha and importin beta. Evaluation of transfection showed higher expression efficiency with plasmid DNA-NLS peptide conjugates than with unmodified plasmids. However, evaluation of intracellular trafficking after microinjection into the cytoplasm showed plasmid DNA-NLS peptide conjugates only within the cytoplasm; there was no NLS-plasmid stimulation of nuclear localization. Our findings suggest that stimulation of plasmid nuclear localization cannot be achieved merely by changing spacer length or chemically modifying plasmid DNA-NLS peptide conjugates. An additional mechanism must be involved.

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Sequence-Specific Photomodification of Single-Stranded and Double-Stranded DNA Fragments by Oligonucleotide Perfluoroarylazide Derivative

Cited by 11 publications

References 18 publications

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

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

Coupling of Nuclear Localization Signals to Plasmid DNA and Specific Interaction of the Conjugates with Importin α

Can Nuclear Localization Signals Enhance Nuclear Localization of Plasmid DNA?

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