In our investigation, employing LC-MS/MS analysis on mitochondrial total RNA, encompassing mt-rRNA, mt-tRNA, and mt-mRNA, we observed a substantial decline in m4C levels, a modest decrease in m5C, and slight reductions in m3C, m7G, and m6A levels following METTL17 knockdown in CRC cells. The gene discussed is METTL17; the disease is colorectal carcinoma.