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Neurosphere cultures transfected with nonsense & miRNA-335 knockout reagents

ABSTRACT: To understand the mechanism by which miR-335 acts anti proliferative factor, we profiled mRNA expression using microarray platform ( codelink array). Embryonic day 12.5 neural cortical epithelium was isolated and grown as neurospheres in defined medium. Cultures were transfected with 2'-O-methyl (2'OMe)-base modified miRNA-335 antisense (2'OMe-335) and control (2' O Me-EGFP). To verify the suppression of miR-335 after the 2'-OMe treatment, we used real time RT-PCR which confirmed the down regulation of miR-335 up to ~4.5 cycles (~24 fold). For the microarray experiment, mRNA isolated from these cells was hybridized to codelink microarrays. Three independent replicate were done for treatment and control, six independent single channel intensity values were uploaded after normalization for analysis. Our array results indicated that there is general up-regulation in gene expression due to miR-335 Knock Out ( KO). We identified 116 genes statistically significant genes (adjusted p value < 0.05), out of these 95.7% were up-regulated genes. The highest fold difference we observed was between 2 and 3 fold, indicating the role of this miRNA in moderately controlling expression hundreds of genes. Of the several genes the highly expressed are important in maturation of neural stem cells and downstream signaling pathway genes

ORGANISM(S): Mus musculus

PROVIDER: GSE14624 | GEO | 2019/07/31

REPOSITORIES: GEO

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Neurosphere cultures transfected with nonsense & miRNA-335 knockout reagents

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