Silencing of mrhl non coding RNA in mouse spermatoginial cells GC1-Spg
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ABSTRACT: Mrhl is a non coding RNA identified from mouse chromosome 8. It is a 2.4kb poly adenylated, nuclear restricted RNA expressed in multiple tissues. The 2.4 kb RNA also undergoes a nuclear processing event mediated through Drosha that generates an 80nt intermediate RNA. This study was aimed at understanding the functiion of mrhl by silencing the mrhl RNA in the mouse spermatogonial cells using a pool of siRNAs targeted against the mrhl and analyse the global gene expression change using Affymetrix mouse expression array. The mRNAs that showed significant change in expression in mrhl siRNA treated cells against control were studied further for their biological significance with respect to mrhl silencing. The mouse spermatogonial cells that were either mock silenced using scrambled siRNA or silenced using siRNAs against mrhl were harvested at 44hrs post transfection. The silencing efficiency was checked by realtime quantitative PCR and was found be almost 80% in all three independent experiments.The total RNA was extracted using Trizol and was taken further for microarray expriment. All the subsequent steps were according to the Affymetrix protocol. Gene expression was profiled for control and mrhl silenced cells.
ORGANISM(S): Mus musculus
SUBMITTER: M Rao
PROVIDER: E-GEOD-19355 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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