Figure 1.

Reprogramming of Dgcr8^Δ/Δ MEFs and TTFs

(A) Schematic of the reprogramming strategy. R26-loxP-STOP-loxP-YFP, ROSA26-driven loxP-flanked STOP sequence followed by an YFP reporter; Ad-Cre, Cre-expressing adenovirus; OSKM, reprogramming factors OCT4, SOX2, KLF4, and c-MYC.

(B) QPCR analyses of mature miRNAs in Dgcr8^flox/flox and Dgcr8^Δ/Δ TTFs 7 or 10 days after Cre expression. Shown are tested miRNAs reliably expressed in Dgcr8^flox/flox TTFs. Expression of mature miRNA was normalized to small nucleolar RNA 142. n = 3 independent biological repeats. Error bar, SD.

(C) Representative flow cytometry analysis of the Dgcr8^Δ/Δ;LoxP-STOP-LoxP-YFP fibroblasts 48 hr after mock (left) or Cre adenovirus (right) transduction. PI, propidium iodide.

(D) Merged bright field and YFP image of fibroblast-derived Dgcr8^Δ/Δ iPSCs. Scale bars, 100 μm.

(E) Reprogramming efficiency of Dgcr8^flox/flox and Dgcr8^Δ/Δ fibroblasts. n = 4 or 5 independent biological repeats. Error bar, SD. See also Table S1.

(F) PCR genotyping of wild-type, Dgcr8^flox/flox TTFs, and Dgcr8^Δ/Δ TTF-derived iPSC clones derived from a representative reprogramming experiment. Although most iPSC clones have Dgcr8 disrupted completely, approximately 15% of YFP+ clones, such as iPSC-5, retain one functional Dgcr8 allele. Diamond, Dgcr8⁺; arrow, Dgcr8^flox; arrowhead, Dgcr8^Δ.