0390087 - ÚMG 2013 RIV US eng J - Journal Article
Nejepínská, Jana - Malík, Radek - Moravec, Martin
Deep sequencing reveals complex spurious transcription from transiently transfected plasmids.
PLoS ONE. Roč. 7, č. 8 (2012), e43283. ISSN 1932-6203. E-ISSN 1932-6203
R&D Projects: GA ČR GA204/09/0085
Grant - others:EMBO(XE) 0001488
Institutional research plan: CEZ:AV0Z50520514
Institutional support: RVO:68378050
Keywords : transient plasmid transfection * deep sequencing
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 3.730, year: 2012

Transient plasmid transfection is a common approach in studies in cultured mammalian cells. To examine behavior of transfected plasmids, we analyzed their transcriptional landscape by deep sequencing. We have found that the entire plasmid sequence is transcribed at different levels. Spurious transcription may have undesirable effects as some plasmids, when co-transfected, inhibited expression of luciferase reporters in a dose-dependent manner. In one case, we attributed this effect to a Kan/Neo resistance cassette, which generated a unique population of edited sense and antisense small RNAs. The unexpected complexity of expression from transiently transfected plasmids underscores the importance of appropriate experimental controls.
Permanent Link: http://hdl.handle.net/11104/0219038