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Statements

Subject Item
n2:RIV%2F00216208%3A11310%2F13%3A10190693%21RIV14-GA0-11310___
rdf:type
skos:Concept n18:Vysledek
rdfs:seeAlso
http://online.liebertpub.com/doi/full/10.1089/dna.2012.1950
dcterms:description
The interferon (IFN) response, induced as a side effect after transfection of nucleic acids into mammalian cells, is known but inadequately described. We followed the IFN response, the fate of cells, and the possible mechanisms leading to this response in NIH3T3 mouse fibroblasts after DNA nucleofection. The gateway destination vector, phGf, and its derivatives encoding toxic and non-toxic variants of the minor structural proteins of polyoma-viruses, VP2 and VP3, were used. DNA vector sequences induced in cells the production of high levels of IFN and the upregulation of the IFN-inducible genes, Mx-1, STAT1, IRF1, and IRF7. The IFN response was not restricted to phGf-derived plasmids. In nucleofected cells, upregulation of the modified gamma-histone 2A.X indicating DNA damage and inhibition of cell proliferation were also observed. Although 3T3 cells expressed the Toll-like receptor-9 (TLR9) and vectors used for nucleofection contained unmethylated CpGs, signaling leading to IFN induction was found to be TLR9 independent. However, the early activation of nuclear factor-kappa B suggested the participation of this transcription factor in IFN induction. Surprisingly, in contrast to nucleofection, transfection using a cationic polymer induced only a poor IFN response. Together, the results point to a strong side effect of nucleofection. The interferon (IFN) response, induced as a side effect after transfection of nucleic acids into mammalian cells, is known but inadequately described. We followed the IFN response, the fate of cells, and the possible mechanisms leading to this response in NIH3T3 mouse fibroblasts after DNA nucleofection. The gateway destination vector, phGf, and its derivatives encoding toxic and non-toxic variants of the minor structural proteins of polyoma-viruses, VP2 and VP3, were used. DNA vector sequences induced in cells the production of high levels of IFN and the upregulation of the IFN-inducible genes, Mx-1, STAT1, IRF1, and IRF7. The IFN response was not restricted to phGf-derived plasmids. In nucleofected cells, upregulation of the modified gamma-histone 2A.X indicating DNA damage and inhibition of cell proliferation were also observed. Although 3T3 cells expressed the Toll-like receptor-9 (TLR9) and vectors used for nucleofection contained unmethylated CpGs, signaling leading to IFN induction was found to be TLR9 independent. However, the early activation of nuclear factor-kappa B suggested the participation of this transcription factor in IFN induction. Surprisingly, in contrast to nucleofection, transfection using a cationic polymer induced only a poor IFN response. Together, the results point to a strong side effect of nucleofection.
dcterms:title
Nucleofection of Expression Vectors Induces a Robust Interferon Response and Inhibition of Cell Proliferation Nucleofection of Expression Vectors Induces a Robust Interferon Response and Inhibition of Cell Proliferation
skos:prefLabel
Nucleofection of Expression Vectors Induces a Robust Interferon Response and Inhibition of Cell Proliferation Nucleofection of Expression Vectors Induces a Robust Interferon Response and Inhibition of Cell Proliferation
skos:notation
RIV/00216208:11310/13:10190693!RIV14-GA0-11310___
n18:predkladatel
n21:orjk%3A11310
n3:aktivita
n12:Z n12:I n12:S n12:P
n3:aktivity
I, P(GAP304/10/1511), S, Z(MSM0021620858)
n3:cisloPeriodika
8
n3:dodaniDat
n4:2014
n3:domaciTvurceVysledku
n9:3219488 n9:5099803 n9:2474174
n3:druhVysledku
n16:J
n3:duvernostUdaju
n15:S
n3:entitaPredkladatele
n7:predkladatel
n3:idSjednocenehoVysledku
92497
n3:idVysledku
RIV/00216208:11310/13:10190693
n3:jazykVysledku
n20:eng
n3:klicovaSlova
microbial dna; dendritic cells; immune-response; mammalian-cells; epithelial-cells; transcription factor; intracellular dna; bacterial-dna; i-interferon; cytosolic dna sensor
n3:klicoveSlovo
n11:epithelial-cells n11:transcription%20factor n11:i-interferon n11:bacterial-dna n11:mammalian-cells n11:intracellular%20dna n11:cytosolic%20dna%20sensor n11:immune-response n11:microbial%20dna n11:dendritic%20cells
n3:kodStatuVydavatele
US - Spojené státy americké
n3:kontrolniKodProRIV
[0B7112795CEB]
n3:nazevZdroje
DNA and Cell Biology
n3:obor
n17:EB
n3:pocetDomacichTvurcuVysledku
3
n3:pocetTvurcuVysledku
3
n3:projekt
n10:GAP304%2F10%2F1511
n3:rokUplatneniVysledku
n4:2013
n3:svazekPeriodika
32
n3:tvurceVysledku
Huérfano- Meneses, Sandra Ryabchenko, Boris Forstová, Jitka
n3:wos
000322315800007
n3:zamer
n13:MSM0021620858
s:issn
1044-5498
s:numberOfPages
13
n19:doi
10.1089/dna.2012.1950
n14:organizacniJednotka
11310