Mutations in the KRAS gene, which persistently activate RAS function, are most frequently found in many types of human
cancers. Here, we proposed and verified a new approach against
cancers harboring the KRAS mutation with high
cancer selectivity and efficient anti-
cancer effects based on targeted
RNA replacement. To this end, trans-splicing
ribozymes from Tetrahymena group I intron were developed, which can specifically target and reprogram the mutant KRAS G12V transcript to induce therapeutic gene activity in cells. Adenoviral vectors containing the specific
ribozymes with downstream suicide gene were constructed and then
infection with the adenoviruses specifically downregulated KRAS G12V expression and killed KRAS G12V-harboring
cancer cells additively upon pro-drug treatment, but it did not affect the growth of wild-type KRAS-expressing cells. Minimal liver toxicity was noted when the adenoviruses were administered systemically in vivo. Importantly, intratumoral injection of the adenoviruses with
pro-drug treatment specifically and significantly impeded the growth of xenografted
tumors harboring KRAS G12V through a trans-splicing reaction with the target
RNA. In contrast, xenografted
tumors harboring wild-type KRAS were not affected by the adenoviruses. Therefore,
RNA replacement with a mutant KRAS-targeting trans-splicing
ribozyme is a potentially useful therapeutic strategy to combat
tumors harboring KRAS mutation.