GLI3 is a zinc finger transcription factor that acts as a dual regulator of sonic hedgehog signaling during development. The full-length GLI3 protein functions as a transcriptional activator, while its C-terminally truncated form (GLI3R) acts as a repressor; a proper balance between these two forms specifies limb digit number and identity. GLI3 also cooperates with TRPS1 to regulate chondrocyte development and binds the consensus sequence 5'-GGGTGGTC-3'. In neural development, GLI3 is required for cortical fate establishment 1 and regulates dorsoventral patterning through direct targets HES4/5. A recent study identified a dual temporal role for GLI3 in neural crest cells, with early GLI3 activity supporting specification and later GLI3 function enabling differentiation into ectomesenchymal derivatives 2. DYRK2-mediated phosphorylation of GLI3 at conserved serine residues promotes its nuclear translocation and activation of hedgehog signaling, particularly during limb development 3. Pathogenic GLI3 variants cause autosomal dominant syndromes including Greig cephalopolysyndactyly syndrome, Pallister-Hall syndrome, and postaxial polydactyly; the mutation location within GLI3 determines phenotype 4. In Pallister-Hall syndrome, truncating GLI3 variants in the middle third of the gene associate with congenital kidney and urinary tract anomalies in 26.9% of reported patients 5. Beyond developmental disorders, GLI3 dysregulation contributes to cancer progression; in gastric cancer, altered GLI3 recruitment enhances USP47 expression to promote epithelial-to-mesenchymal transition 6.