Engineered targeted enhancement of translation in mammalian cell

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2026年6月13日

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Junzhe Liu,
Yonghui Lv,
Zhihong Zhou,
Linzhen Huang,
Kunjian Lei,
Zewei Tu,
Chuandong Gong,
Zhansheng Fang,
Chong Wang,
Juan Wang,
Xingen Zhu,
Minhua Ye,
Shigang Lv,
Kai Huang

摘要

Background

While the central dogma outlines DNA-to-protein information flow, existing gene regulators mainly target transcription. Here, we developed the λN-Guided RNA Targeting System (λGRTS), a CRISPR-independent platform enhancing mammalian mRNA translation via specific translation-guiding RNAs (tgRNAs).

Results

λGRTS integrates λN (high-affinity BoxB binder), mutated eIF4E1 (ablated non-specific 5′ cap binding, retains TIC recruitment), and auxiliary factors (HuR for dsRNA stabilization, PABP and RRM2-RRM3 for mRNA closed loops). Optimized 22-nt tgRNAs (targeting 58 bp upstream of mRNA ATG in 5′ UTR) and NES-tagged proteins maximized efficacy. λGRTS outperformed dCasRX (smaller ~ 50 kDa vs. ~ 150 kDa, single tgRNA vs. crRNA-tracrRNA), boosting functional proteins (e.g., GFP). It activated P53/PTEN, suppressing GBM cell proliferation, inducing G1 arrest, and reducing invasion in vitro. In vivo, lentiviral λGRTS inhibited orthotopic GBM in nude mice, extended survival by approximately 35–40 days, with IHC confirming P53/PTEN upregulation. Mass spectrometry showed no off-target effects. Cross-species tests (human, mouse, bovine cells) validated broad applicability via conserved eIF4E1.

Conclusions

λGRTS offers a specific, safe translation-centric tool for gene regulation and oncology research.

Graphical Abstract