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| Status |
Public on Aug 17, 2012 |
| Title |
Complexity of expression from transiently transfected plasmids |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Transient plasmid transfection is common approach for studies in cultured mammalian cells. To examine behavior of transfected plasmids, we analyzed their transcriptional landscape by deep sequencing. We found that plasmids generate different levels of transcripts virtually everywhere. Spurious transcription may have undesirable effects as some co-transfected plasmids inhibited expression of luciferase reporters in a dose-dependent manner. In one case, we attributed this effect to kan/neo resistance cassette, which generated a unique population of edited sense and antisense small RNAs. The unexpected complexity of expression of transiently transfected plasmids highlights the importance of appropriate experimental controls.
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| Overall design |
HEK293 cells (human origin) transiently transfected with 4 various plasmids
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| Contributor(s) |
Nejepinska J, Malik R, Moravec M, Svoboda P |
| Citation(s) |
22916237 |
| Submission date |
Feb 24, 2012 |
| Last update date |
May 15, 2019 |
| Contact name |
Radek Malik |
| E-mail(s) |
malikr@img.cas.cz
|
| Organization name |
Institute of Molecular Genetics of the ASCR
|
| Department |
Dep. of Epigenetics Regulations
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| Street address |
Videnska 1083
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| City |
Prague 4 |
| ZIP/Postal code |
CZ-14220 |
| Country |
Czech Republic |
| |
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| Platforms (1) |
| GPL13393 |
AB SOLiD 4 System (Homo sapiens) |
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| Samples (3) |
| GSM880643 |
HEK293 cells transfected with phRL-SV40 plasmid |
| GSM880644 |
HEK293 cells transfected with pGL4-SV40 plasmid |
| GSM880645 |
HEK293 cells transfected with a 1:1 mixture of pBluescript and pEGFP-C1 plasmids |
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| Relations |
| SRA |
SRP011043 |
| BioProject |
PRJNA151813 |
