Engineered targeted enhancement of translation in mammalian cell.

IF 5.4 1区生物学 Q1 BIOLOGY

BMC Biology Pub Date : 2026-06-13 DOI:10.1186/s12915-026-02653-0

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

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引用次数: 0

Abstract

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.

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在哺乳动物细胞中工程化靶向增强翻译。

背景：虽然中心法则概述了dna到蛋白质的信息流，但现有的基因调控主要针对转录。在这里，我们开发了λN-Guided RNA Targeting System (λGRTS)，这是一个不依赖crispr的平台，通过特异性翻译引导RNA （tgRNAs）增强哺乳动物mRNA的翻译。结果：λGRTS整合了λN（高亲和力BoxB结合剂）、突变的eIF4E1（非特异性5'帽结合，保留TIC招募）和辅助因子（用于dsRNA稳定的HuR、用于mRNA闭合环的PABP和RRM2-RRM3）。优化的22-nt tgRNAs（靶向5' UTR中mRNA ATG上游58 bp）和nes标记蛋白的效果最大。λGRTS优于dCasRX（较小~ 50 kDa vs ~ 150 kDa，单个tgRNA vs crRNA-tracrRNA），促进功能蛋白（如GFP）。激活P53/PTEN，抑制GBM细胞增殖，诱导G1阻滞，减少体外侵袭。在体内，慢病毒λGRTS抑制裸小鼠原位GBM，延长存活时间约35-40天，IHC证实P53/PTEN上调。质谱分析未发现脱靶效应。跨物种测试（人类、小鼠、牛细胞）证实了保守的eIF4E1的广泛适用性。结论：λGRTS为基因调控和肿瘤研究提供了一个特定的、安全的以翻译为中心的工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

BMC Biology 生物-生物学

CiteScore

7.80

自引率

1.90%

发文量

260

审稿时长

3 months

期刊介绍： BMC Biology is a broad scope journal covering all areas of biology. Our content includes research articles, new methods and tools. BMC Biology also publishes reviews, Q&A, and commentaries.