近日，德国合成生物医学研究所教授Dong-Jiunn Jeffery Truong及其团队开发出用于装载可编程编辑器的工程核细胞质载具。相关论文于2025年4月9日发表于国际顶尖学术期刊《细胞》杂志上。

在这项研究中，研究团队提出了一个病毒样粒子（VLP）系统，该系统具有核细胞质穿梭载体，通过适体标记的引导RNA检索预组装的Cas效应物。这种方法确保了完全组装的编辑核糖核蛋白（RNPs）的优先装载，并提高了在多种永生化、原代、干细胞和干细胞衍生细胞类型中，与同源定向修复相关的引物编辑、碱基编辑、反式激活剂和核酸酶活性的功效。小组还通过用Csy4/Cas6f屏蔽3 '' -暴露端，实现了对固有不稳定的引物编辑引导RNA（pegRNAs）的额外保护，进一步提高了编辑性能。

此外，课题组研究人员确定了一套最小的包装和萌芽模块，可以作为一个平台，自下而上的工程包裹投递车辆。值得注意的是，他们的系统在原发性T淋巴细胞和遗传性视网膜疾病的两种小鼠模型中显示出优越的per-VLP编辑效率，突出了其治疗潜力。

据了解，先进的基因编辑方法加速了生物医学的发现，并具有巨大的治疗前景，但安全有效地传递基因编辑器仍然具有挑战性。

附：英文原文

Title: Engineered nucleocytosolic vehicles for loading of programmable editors

Author: Julian Geilenkeuser, Niklas Armbrust, Emily Steinmal, Samuel W. Du, Sebastian Schmidt, Eva Maria Hildegard Binder, Yuchun Li, Niklas Wilhelm Warsing, Stephanie Victoria Wendel, Florian von der Linde, Elisa Marie Schiele, Xiya Niu, Luisa Stroppel, Oleksandr Berezin, Tobias Heinrich Santl, Tanja Orschmann, Keith Nelson, Christoph Gruber, Grazyna Palczewska, Carolline Rodrigues Menezes, Eleonora Risaliti, Zachary J. Engfer, Naile Koleci, Andrea Schmidts, Arie Geerlof, Krzysztof Palczewski, Gil Gregor Westmeyer, Dong-Jiunn Jeffery Truong

Issue&Volume: 2025-04-09

Abstract: Advanced gene editing methods have accelerated biomedical discovery and hold great therapeutic promise, but safe and efficient delivery of gene editors remains challenging. In this study, we present a virus-like particle (VLP) system featuring nucleocytosolic shuttling vehicles that retrieve pre-assembled Cas-effectors via aptamer-tagged guide RNAs. This approach ensures preferential loading of fully assembled editor ribonucleoproteins (RNPs) and enhances the efficacy of prime editing, base editing, trans-activators, and nuclease activity coupled to homology-directed repair in multiple immortalized, primary, stem cell, and stem-cell-derived cell types. We also achieve additional protection of inherently unstable prime editing guide RNAs (pegRNAs) by shielding the 3′-exposed end with Csy4/Cas6f, further enhancing editing performance. Furthermore, we identify a minimal set of packaging and budding modules that can serve as a platform for bottom-up engineering of enveloped delivery vehicles. Notably, our system demonstrates superior per-VLP editing efficiency in primary T lymphocytes and two mouse models of inherited retinal disease, highlighting its therapeutic potential.

DOI: 10.1016/j.cell.2025.03.015

Source: https://www.cell.com/cell/abstract/S0092-8674(25)00288-0