DDX50 shRNA knock down or control shRNA in skin organoids from primary neonatal keratinocytes to determine the function of DDX50 in skin homeostasis.
Primary human keratinocytes were isolated from fresh surgically discarded newborn foreskin, and progenitor cells cultured in complete Keratinocyte-SFM (Life Technologies, #17005-142) and Medium 154 (Life Technologies #M-154-500). Organotypic regeneration of human epidermal and cervical tissues were performed as previously described (https://doi.org/10.1101/gad.1463206) using KGM media. Day 7 epidermal and cervical tissues were collected. Total RNA was extracted using the RNeasy Plus kit (QIAGEN, 74136). QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen, 015.96)
NIAMS-phs003766
a_shDDX50-4_biorep4_techrep1_organoid