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

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.

Species:

mouse

Samples:

10

Source:

E-MEXP-1455

PubMed:

18329371

Updated:

Dec.12, 2014

Registered:

Nov.21, 2014