CRISPR/Cas9 mediated knock-out of RUNX2 in melanoma cells

Incidence of MM has increased considerably in consequence of lifestyle and environmental changes. The mortality rate for MM is very high as it is highly invasive and also genetically resistant to chemotherapeutic treatments. It has been reported that mutation rate and gene modulation in melanoma are higher than in other solid malignancies. In addition, transcription factors by acting on gene expression can affect cellular process. In particular a higher expression of RUNX2 in melanoma than in normal melanocytes have been shown. RUNX2 is overexpressed in several tumor tissues, including pancreatic cancer , breast cancer, ovarian epithelial cancer , prostate cancer , lung cancer and osteosarcoma. As no direct RUNX2 inhibitor is available and experiments performed with RNA interference were scarcely reproducible we applied Crisp/cas 9 technology to knockout the RUNT domain of RUNX2 in melanoma cell line. Crisp/Cas 9 tecnology was able to delete, partially, the RUNT domain. The deleted clone showed a reduced proliferation, reduced EMT features, reduced migration ability, suggesting the involvement of RUNT in different pathway of MM. In addition, the deleted clone showed a reduction of genes involved in migration ability and an increased expression of SSBP1 gene suggesting RUNT as oncotarget in MM.