|
| Status |
Public on Dec 20, 2012 |
| Title |
Airn transcriptional overlap but not its lncRNA products induce imprinted Igf2r silencing. |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by genome tiling array
|
| Summary |
To test if the imprinted long non-coding RNA (lncRNA) Airn transcription or its product silences the protein-coding Igf2r gene, we shortened the endogenous lncRNA to four different lengths by inserting polyadenylation (polyA) cassettes on the paternal chromosome via homologous recombination in ES cells.
ES cell differentiation was used to recapitulate the developmental onset of Airn and Igf2r imprinted expression and polyA cassettes inserted before (T3, T16, T27) or after (T31, T51) the Igf2r promoter successfully truncated Airn, with the exception of T27.
RNA hybridization to a tiling array (MIRTA) demonstrated loss of Airn upstream of the Igf2r promoter (except T27) and absence of novel spliced variants in all truncation alleles.
To further test the transcriptional overlap model we moved the Airn promoter ~700bp before the Igf2r TSS in ES cells that lack an endogenous paternal Airn promoter and called this cell line FAP (Forward-Airn-Promoter).
RNA hybridization to a tiling array (MIRTA) demonstrated that the moved Airn promoter expresses Airn in wildtype orientation and terminates at the wildtype 3' end.
|
| |
|
| Overall design |
ES cell lines expressing different Airn variants were differentiated on gelatinized dishes after feeder cell depletion and LIF withdrawal by 0.27µM retinoic acid for 5 days and Airn length was assayed.
|
| |
|
| Contributor(s) |
Latos PA, Pauler FM, Koerner MV, Şenergin HB, Barlow DP |
| Citation(s) |
23239737 |
| |
| Submission date |
Oct 10, 2012 |
| Last update date |
Dec 22, 2012 |
| Contact name |
Florian M Pauler |
| E-mail(s) |
florian.pauler@ist.ac.at
|
| Phone |
+43 2243 9000-7434
|
| Organization name |
IST Austria
|
| Lab |
Simon Hippenmeyer
|
| Street address |
Am Campus 1
|
| City |
Klosterneuburg |
| ZIP/Postal code |
3400 |
| Country |
Austria |
| |
|
| Platforms (1) |
| GPL11618 |
NimbleGen custom Mouse MIRTA Array |
|
| Samples (7)
|
|
| Relations |
| BioProject |
PRJNA177027 |
Less...
More...