Wnt signaling has emerged as a key regulator of maintaining gut epithelial renewal and homeostasis. Wnt proteins locally interact with and activate the Frizzled (Fz) receptors, leading to stimulation of their target gene transcription by increasing β-catenin nuclear translocation. Our recent study shows that activation of Wnt3a/β-catenin signaling enhances intestinal epithelial repair by promoting c-Myc-regulated gene expression, but the exact mechanism that regulates Fz-receptor abundance remains unknown. microRNAs (miRNAs) and RNA-binding proteins (RBPs) regulate gene expression at the posttranscriptional level. miRNA-222 (miR-222) and RBP CUG-binding protein 1 (CUGBP1) are highly expressed in the gut mucosa and they are involved in many aspects of cellular function In Vitro and In Vivo. In this study, we test the hypothesis that miR-222 and CUGBP1 jointly regulate Fz7 expression in intestinal epithelial cells (IECs) and therefore the epithelial repair after injury. Methods: Studies were conducted in IEC-6 cells, which were derived from rat intestinal crypts. Interaction of CUGBP1 with the Fz-7 mRNA was detected by biotin pull-down and RNP-IP assays, whereas miR-222 binding to the Fz-7 transcript was examined by RNA pulldown assay using biotin-labeled miR-222 and pmir-Glo reporter system. Fz-7 translation was examined by measuring its newly synthesized protein and using chimeric luciferaseFz-7 coding region (CR) and 3'-untranslated region (UTR) reporter gene assays. CUGBP1 andmiR-222 functions were investigated by siRNA silencing and ectopic gene overexpression. Epithelial repair was measured in an In Vitro wounding model. Results: miR-222 was found to bind the Fz-7 mRNA via the CR rather than its 5'and 3'-UTRs, whereas CUGBP1 bound to both 3'-UTR and CR of the Fz-7 mRNA. Overexpression of a precursor miR-222 increased miR-222/Fz-7 mRNA complex, repressed Fz-7 translation, and reduced its protein levels (by ~60%), although it failed to alter total Fz-7 mRNA levels. miR-222 silencing decreased miR-222 association with the Fz-7 mRNA, enhanced Fz-7 translation, and increased its protein content (by ~2-fold). Ectopic CUGBP1 overexpression also repressed Fz-7 translation without effect on Fz-7 mRNA levels, while CUGBP1 silencing stimulated Fz-7 expression. Interestingly, co-transfection of the miR-222 precursor with CUGBP1 synergistically repressed Fz-7 translation; CUGBP1 silencing partially abolished the Fz-7 repression in cells overexpressing miR-222. Ectopic overexpression of CUGBP1 and miR-222 or Fz-7 silencing repressed Wnt-dependent transcriptional activity and suppressed epithelial repair after wounding Conclusions: These results indicate that CUGBP1 and miR-222 synergistically repress Fz-7 translation and inactivate Wnt signaling pathway, thus inhibiting epithelial repair after wounding.