美国哈佛大学David R. Liu等研究人员合作定向演化工程病毒样颗粒来提高生产和转导效率。该研究于2024年11月13日在线发表于国际一流学术期刊《自然—生物技术》。

研究人员报道了一种用于实验室演化工程病毒样颗粒（eVLP）的系统，该系统能够发现具有改进特性的eVLP变体。该系统通过在DNA自由的eVLP包装货物中加载带条形码的导向RNA，唯一标记每个eVLP变体，从而在筛选期望特性后识别出所需的变体。研究人员将此系统应用于突变和筛选具有改进的eVLP生产特性或在人类细胞中具有更高转导效率的eVLP外壳蛋白。

通过结合有益的外壳蛋白突变，研究人员开发了第五代（v5）eVLP，这些颗粒在培养的哺乳动物细胞中的递送效力比之前的最佳v4 eVLP提高了2-4倍。对v5 eVLP的分析表明，这些外壳蛋白突变优化了所需核糖核蛋白货物的包装和递送，而非原生病毒基因组，并显著改变了eVLP外壳蛋白的结构。这些发现表明，带条形码的eVLP演化有潜力支持更高效eVLP的开发。

据了解，eVLP是用于瞬时递送蛋白质和RNA（包括基因编辑工具）的有前景的载体。

附：英文原文

Title: Directed evolution of engineered virus-like particles with improved production and transduction efficiencies

Author: Raguram, Aditya, An, Meirui, Chen, Paul Z., Liu, David R.

Issue&Volume: 2024-11-13

Abstract: Engineered virus-like particles (eVLPs) are promising vehicles for transient delivery of proteins and RNAs, including gene editing agents. We report a system for the laboratory evolution of eVLPs that enables the discovery of eVLP variants with improved properties. The system uses barcoded guide RNAs loaded within DNA-free eVLP-packaged cargos to uniquely label each eVLP variant in a library, enabling the identification of desired variants following selections for desired properties. We applied this system to mutate and select eVLP capsids with improved eVLP production properties or transduction efficiencies in human cells. By combining beneficial capsid mutations, we developed fifth-generation (v5) eVLPs, which exhibit a 2–4-fold increase in cultured mammalian cell delivery potency compared to previous-best v4 eVLPs. Analyses of v5 eVLPs suggest that these capsid mutations optimize packaging and delivery of desired ribonucleoprotein cargos rather than native viral genomes and substantially alter eVLP capsid structure. These findings suggest the potential of barcoded eVLP evolution to support the development of improved eVLPs.

DOI: 10.1038/s41587-024-02467-x

Source: https://www.nature.com/articles/s41587-024-02467-x