Programmable RNA-triggered cancer cell elimination using CRISPR-Cas12a2

Selective eradication of cancer without harming healthy tissue is vital to precision oncology. Although traditional interventions such as chemotherapy or ionizing radiation are broadly applicable across many cancer types, they often exhibit off-target cytotoxicity; conversely, targeted small molecules and biologics improve specificity by binding to protein mutations, yet may not be as broadly applicable. Developing a therapeutic approach with both high specificity and broad applicability thus remains a major challenge in modern cancer care. Here, we show that Cas12a2, a recently discovered CRISPR nuclease exhibiting RNA-triggered DNA shredding, enables programmable sequence-specific elimination of mammalian cells expressing a target transcript. Target-expressing cultures electroporated with NLS-tagged Cas12a2 undergo massive cell death within five days. Activating Cas12a2 elicits widespread double-strand DNA breaks in the nucleus, leading to mitotic catastrophe, cellular inflammation, and apoptosis. We demonstrate that Cas12a2 distinguishes different RNA targets within heterogeneous cultures and induces cell death without observable off-target activity. Leveraging this approach, we eliminate NCI-H23 cancer cells harboring the prevalent oncogenic KRAS (G12C) point mutation, including cells with resistance to the FDA-approved KRAS (G12C) inhibitor Sotorasib. Taken together, these findings present Cas12a2 as a specific and broadly applicable therapeutic platform for personalized cancer treatment. These findings further establish the basis for use of Cas12a2 as a potent cell ablation tool across disciplines of basic and applied research.