scholarly journals Exhaustivein vivolabelling of plasmid DNA with BrdU for intracellular detection in non-viral transfection of mammalian cells

2009 ◽  
Vol 4 (10) ◽  
pp. 1479-1487 ◽  
Author(s):  
Valérie Jérôme ◽  
Andreas Heider ◽  
Anja Schallon ◽  
Ruth Freitag
Keyword(s):  
Plasmid Dna ◽  
Mammalian Cells ◽  
Intracellular Detection ◽  
Viral Transfection

Hamster polyomavirus-derived virus-like particles are able to transfer in vitro encapsidated plasmid DNA to mammalian cells

Virus Genes ◽  
2007 ◽  
Vol 34 (3) ◽  
pp. 303-314 ◽  
Author(s):  
Tatyana Voronkova ◽  
Andris Kazaks ◽  
Velta Ose ◽  
Muhsin Özel ◽  
Siegfried Scherneck ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Mammalian Cells ◽  
Virus Like Particles ◽  
Hamster Polyomavirus

scholarly journals Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian cell nuclei.

1985 ◽  
Vol 5 (1) ◽  
pp. 59-69 ◽  
Author(s):  
K R Folger ◽  
K Thomas ◽  
M R Capecchi
Keyword(s):  
Homologous Recombination ◽  
Dna Sequences ◽  
Plasmid Dna ◽  
Mammalian Cells ◽  
Recombinant Dna ◽  
Dna Duplexes ◽  
Dna Molecules ◽  
Cell Nuclei ◽  
High Concentration ◽  
Length Polymorphisms

We have examined the mechanism of homologous recombination between plasmid molecules coinjected into cultured mammalian cells. Cell lines containing recombinant DNA molecules were obtained by selecting for the reconstruction of a functional Neor gene from two plasmids that bear different amber mutations in the Neor gene. In addition, these plasmids contain restriction-length polymorphisms within and near the Neor gene. These polymorphisms did not confer a selectable phenotype but were used to identify and categorize selected and nonselected recombinant DNA molecules. The striking conclusion from this analysis is that the predominant mechanism for the exchange of information between coinjected plasmid molecules over short distances (i.e., less than 1 kilobase) proceeds via nonreciprocal homologous recombination. The frequency of homologous recombination between coinjected plasmid molecules in cultured mammalian cells is extremely high, approaching unity. We demonstrate that this high frequency requires neither a high input of plasmid molecules per cell nor a localized high concentration of plasmid DNA within the nucleus. Thus, it appears that plasmid molecules, once introduced into the nucleus, have no difficulty seeking each other out and participating in homologous recombination even in the presence of a vast excess of host DNA sequences. Finally, we show that most of the homologous recombination events occur within a 1-h interval after the introduction of plasmid DNA into the cell nucleus.

Amphiphilic azobenzene-neomycin conjugate self-assembles into nanostructures and transports plasmid DNA efficiently into the mammalian cells

2016 ◽  
Vol 148 ◽  
pp. 481-486 ◽  
Author(s):  
Santosh Yadav ◽  
Smriti Rekha Deka ◽  
Diksha Jha ◽  
Hemant Kumar Gautam ◽  
Ashwani Kumar Sharma
Keyword(s):  
Plasmid Dna ◽  
Mammalian Cells

Electroporation-mediated transfection of mammalian cells with crude plasmid DNA preparations

1995 ◽  
Vol 12 (2) ◽  
pp. 113-117 ◽  
Author(s):  
Masaaki Tatsuka ◽  
Nobuyuki Yamagishi ◽  
Morimasa Wada ◽  
Hiromi Mitsui ◽  
Takahide Ota ◽  
...  
Keyword(s):  
Plasmid Dna ◽  
Mammalian Cells

scholarly journals The study of complexation between dicationic surfactants and the DNA duplex using structural and spectroscopic methods

RSC Advances ◽  
2017 ◽  
Vol 7 (42) ◽  
pp. 26006-26018 ◽  
Author(s):  
W. Andrzejewska ◽  
M. Wilkowska ◽  
M. Chrabąszczewska ◽  
M. Kozak
Keyword(s):  
Nucleic Acids ◽  
Plasmid Dna ◽  
Living Cells ◽  
Spectroscopic Methods ◽  
Dna Duplex ◽  
Dna Oligomers ◽  
Different Types ◽  
Dicationic Surfactants ◽  
Viral Transfection

Dicationic (also known as gemini or dimeric) bis-alkylimidazolium surfactants belong to a group of non-viral transfection systems proposed for the successful introduction of different types of nucleic acids (i.e., siRNA, DNA oligomers, and plasmid DNA) into living cells.

scholarly journals Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

2021 ◽  
Vol 9 ◽  
Author(s):  
James D. Budge ◽  
Robert J. Young ◽  
Christopher Mark Smales
Keyword(s):  
Recombinant Protein ◽  
Plasmid Dna ◽  
Cho Cells ◽  
Mammalian Cells ◽  
Chinese Hamster Ovary Cells ◽  
Transient Gene Expression ◽  
Chinese Hamster ◽  
Nuclear Antigen ◽  
Stable Cell Line ◽  
Nuclear Pore

Transient gene expression (TGE) in mammalian cells is a method of rapidly generating recombinant protein material for initial characterisation studies that does not require time-consuming processes associated with stable cell line construction. High TGE yields are heavily dependent on efficient delivery of plasmid DNA across both the plasma and nuclear membranes. Here, we harness the protein nucleoside diphosphate kinase (NDPK-A) that contains a nuclear localisation signal (NLS) to enhance DNA delivery into the nucleus of CHO cells. We show that co-expression of NDPK-A during transient expression results in improved transfection efficiency in CHO cells, presumably due to enhanced transportation of plasmid DNA into the nucleus via the nuclear pore complex. Furthermore, introduction of the Epstein Barr Nuclear Antigen-1 (EBNA-1), a protein that is capable of inducing extrachromosomal maintenance, when coupled with complementary oriP elements on a transient plasmid, was utilised to reduce the effect of plasmid dilution. Whilst there was attenuated growth upon introduction of the EBNA-1 system into CHO cells, when both NDPK-A nuclear import and EBNA-1 mediated technologies were employed together this resulted in enhanced transient recombinant protein yields superior to those generated using either approach independently, including when expressing the complex SARS-CoV-2 spike (S) glycoprotein.

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