Engineered pistol ribozymes selectively target KRAS G12V with enhanced efficacy by capping modification

Zhenzhen Li¹, Ming Zhao², Zhiqin Xi², Jie Bai², Ying Zhang³, Xuelin Zhan⁴, Yili Yang⁵, Yijin Liu⁶

Affiliations

¹ State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Haihe Education Park, No. 38 Tongyan Road, Tianjin 300071, P.R. China; China Regional Research Centre, International Centre for Genetic Engineering and Biotechnology, 8 Taohongjing Road, Taizhou, Jiangsu 225316, P.R. China.
² State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Haihe Education Park, No. 38 Tongyan Road, Tianjin 300071, P.R. China.
³ School of Medicine, University of St Andrews, North Haugh, St Andrews, Fife KY16 9TF, UK.
⁴ China Regional Research Centre, International Centre for Genetic Engineering and Biotechnology, 8 Taohongjing Road, Taizhou, Jiangsu 225316, P.R. China.
⁵ China Regional Research Centre, International Centre for Genetic Engineering and Biotechnology, 8 Taohongjing Road, Taizhou, Jiangsu 225316, P.R. China. Electronic address: yili.yang@icgeb.cn.
⁶ State Key Laboratory of Medicinal Chemical Biology, Frontiers Science Center for New Organic Matter, Frontiers Science Center for Cell Responses and College of Pharmacy, Nankai University, Haihe Education Park, No. 38 Tongyan Road, Tianjin 300071, P.R. China. Electronic address: yvliu@nankai.edu.cn.

PMID: 41864209
DOI: 10.1016/j.chembiol.2026.03.001

Abstract

In this study, we rationally engineered a previously characterized pistol ribozyme to selectively target the KRAS p.G12V mutation with high specificity and efficiency. The pseudoknot-induced compact folding structure of the pistol ribozyme provides three-dimensional structural recognition of the KRAS substrate, distinguishing it from siRNA and ASO, which rely solely on primary sequence complementarity. This structural advantage enables the designed pistol ribozyme to effectively differentiate between KRAS wild-type and G12V mutant RNA. Furthermore, compared with 3′ end nucleotide derivative modifications, 5′ end capping is more effective at increasing ribozyme stability and compatibility with in vitro preparations. These features underscore the promising potential of natural pistol ribozymes as advanced therapeutic nucleic acid molecules for targeting KRAS mutation-driven cancers and suggest a generalizable strategy for structure-guided, allele-specific RNA therapeutics.

Keywords: KRAS mutation; catalysis; gene therapy; modification; pistol ribozyme.

Cell Chem Biol. 2026 Mar 20:S2451-9456(26)00071-1.doi: 10.1016/j.chembiol.2026.03.001. (IF:7.2).

Engineered pistol ribozymes selectively target KRAS G12V with enhanced efficacy by capping modification

Abstract

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