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Transcription profiling of mouse Abl pre-B cell lines with conditional deletion of dicer

ABSTRACT: Abelson virus (v-Abl)-transformed pre-B cell lines from BM of dcrfl/fl; R26R-yfp (4470; 4475) and dcrfl/+; R26R-yfp mice (4483) are treated with a transducible Cre protein (Tat-Cre) to induce deletion of the dicer-1 gene in vitro. Cre activity is conveniently monitored by concomitant expression of YFP. To obtain Dicer KO cells, YFP+ cells from dcrfl/fl; R26R-yfp (4470_YFP; 4475_YFP) cultures treated with Tat-cre were isolated by fluorescence-activated cell sorting (FACS) and further propagated in cell culture. As controls, we used non-deleted, YFP- cells sorted from dcrfl/fl; R26R-yfp cultures (4470; 4475) treated with Tat-cre, and YFP+ deleted cells from dcrfl/+; R26R-yfp cultures (4483_YFP). To validate this system, various cell populations were sorted and analyzed for Dicer protein and miRNA expression. In the Dicer KO pro-B cell lines, Western analysis indicated that Dicer protein was efficiently ablated, and Northern blots demonstrated that the levels of mature miR-17 and miR-191 were drastically reduced.

INSTRUMENT(S): 418 [Affymetrix]

ORGANISM(S): Mus musculus  

SUBMITTER: Gunther R Galler  

PROVIDER: E-MEXP-1455 | ArrayExpress | 2008-03-08


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To explore the role of Dicer-dependent control mechanisms in B lymphocyte development, we ablated this enzyme in early B cell progenitors. This resulted in a developmental block at the pro- to pre-B cell transition. Gene-expression profiling revealed a miR-17 approximately 92 signature in the 3'UTRs of genes upregulated in Dicer-deficient pro-B cells; a top miR-17 approximately 92 target, the proapoptotic molecule Bim, was highly upregulated. Accordingly, B cell development could be partially re  ...[more]

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