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Mammary gland development is fueled by stem cell self‐renewal and differentiation. External cues from the microenvironment coupled with internal cues such as post‐transcriptional regulation exerted by microRNAs regulate stem cell behavior and fate. Here, we have identified a miR‐205 regulatory network required for mammary gland ductal development and stem cell regeneration following transplantation into the cleared mammary fat pad. In the postnatal mammary gland, miR‐205 is predominantly expressed in the basal/stem cell enriched population. Conditional deletion of miR‐205 in mammary epithelial cells impairs stem cell self‐renewal and mammary regenerative potential in the in vitro mammosphere formation assay and in vivo mammary reconstitution. miR‐205 null transplants display significant changes in basal cells, basement membrane, and stroma. NKD1 and PTPA, which inhibit the Wnt signaling pathway, and AMOT, which causes YAP cytoplasmic retention and inactivation were identified as miR‐205 downstream mediators. These studies also confirmed that miR‐205 is a direct Δ Np63  target gene that is critical for the regulation of basal cell identity.  Stem Cells   2018;36:1875–15

miR‐205 Regulates Basal Cell Identity and Stem Cell Regenerative Potential During Mammary Reconstitution