Miniature and versatile genome regulation TnpB-&#x3c9;RNA toolkits facilitate cancer immunotherapy.

Junlin Lu; Jiajian Lai; Liang Cheng; Hengji Zhan; Kaiwen Jie; Chu Liu; Lin Huang; Meifeng Cen; Sen Liu; Zehua Chen; Qiang Zhang; Jianhui Zhang; Jingdie Wu; Bolin Pan; Siting Chen; Jianmei Zhong; Baoqing He; Hui Li; Xu Chen; Tianxin Lin

doi:10.1038/s41467-026-71327-w

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PubMed · 41917051

Miniature and versatile genome regulation TnpB-ωRNA toolkits facilitate cancer immunotherapy.

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Abstract

CRISPR‒Cas systems represent powerful tools for genome regulation. However, the large size of Cas proteins limits their efficient delivery via an adeno-associated virus (AAV), thereby restricting their clinical translation. Here, we engineer the IS200/IS605 transposon-encoded nuclease TnpB, along with its ωRNA scaffold, to create an enhanced TnpB system, which serves as a compact toolkit for gene activation, genome editing, and base editing. The gene activator enTnpBa increases expression by 2889-fold with a minimized 93 nt ωRNA and robustly activates endogenous genes in mammalian cells. We develop a single-AAV-based regimen for immune activation (AAV-ImmunAct) that delivers enTnpBa to activate CXCL9, IL-15, and IFN-γ. AAV-ImmunAct effectively enhances T cell migration and activation, increases killing of cancer cell lines and patient-derived organoids, and synergizes with anti-PD-1 therapy in humanized mice. Here, we establish enTnpB as a compact and versatile platform for genome regulation and a promising tool for cancer immunotherapy.

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Humans Animals Immunotherapy CRISPR-Cas Systems Mice Cell Line, Tumor Dependovirus Neoplasms Gene Editing T-Lymphocytes Journal Article

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Junlin Lu, Jiajian Lai, Liang Cheng, Hengji Zhan, Kaiwen Jie, Chu Liu, Lin Huang, Meifeng Cen, Sen Liu, Zehua Chen, Qiang Zhang, Jianhui Zhang, Jingdie Wu, Bolin Pan, Siting Chen, Jianmei Zhong, Baoqing He, Hui Li, Xu Chen, Tianxin Lin. 2026-04-01. Miniature and versatile genome regulation TnpB-ωRNA toolkits facilitate cancer immunotherapy.. https://doi.org/10.1038/s41467-026-71327-w

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