• Something wrong with this record ?

Deep sequencing reveals complex spurious transcription from transiently transfected plasmids

J. Nejepinska, R. Malik, M. Moravec, P. Svoboda,

. 2012 ; 7 (8) : e43283.

Language English Country United States

Document type Journal Article, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc13012517

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.

References provided by Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc13012517
003      
CZ-PrNML
005      
20130408112424.0
007      
ta
008      
130404s2012 xxu f 000 0|eng||
009      
AR
024    7_
$a 10.1371/journal.pone.0043283 $2 doi
035    __
$a (PubMed)22916237
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxu
100    1_
$a Nejepinska, Jana $u Institute of Molecular Genetics AS CR, Prague, Czech Republic.
245    10
$a Deep sequencing reveals complex spurious transcription from transiently transfected plasmids / $c J. Nejepinska, R. Malik, M. Moravec, P. Svoboda,
520    9_
$a 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.
650    _2
$a buněčné linie $7 D002460
650    _2
$a průtoková cytometrie $7 D005434
650    _2
$a vysoce účinné nukleotidové sekvenování $x metody $7 D059014
650    _2
$a lidé $7 D006801
650    _2
$a plazmidy $x genetika $7 D010957
650    _2
$a antisense RNA $x genetika $7 D016372
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Malik, Radek $u -
700    1_
$a Moravec, Martin $u -
700    1_
$a Svoboda, Petr $u -
773    0_
$w MED00180950 $t PloS one $x 1932-6203 $g Roč. 7, č. 8 (2012), s. e43283
856    41
$u https://pubmed.ncbi.nlm.nih.gov/22916237 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20130404 $b ABA008
991    __
$a 20130408112651 $b ABA008
999    __
$a ok $b bmc $g 975715 $s 810798
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2012 $b 7 $c 8 $d e43283 $i 1932-6203 $m PLoS One $n PLoS One $x MED00180950
LZP    __
$a Pubmed-20130404

Find record

Citation metrics

Archiving options