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

IF 33.1 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Aditya Raguram, Meirui An, Paul Z. Chen, David R. Liu
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引用次数: 0

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.

Abstract Image

改进生产和转导效率的工程病毒颗粒的定向进化
工程病毒样颗粒(eVLPs)是一种很有前景的蛋白质和 RNA(包括基因编辑剂)瞬时递送载体。我们报告了一种用于 eVLP 实验室进化的系统,它能发现具有更好特性的 eVLP 变体。该系统利用装载在无 DNA eVLP 包装货物中的条形码引导 RNA 来唯一标记文库中的每个 eVLP 变体,从而能够在根据所需的特性进行选择后识别出所需的变体。我们应用该系统对 eVLP 胶囊进行了突变和筛选,从而提高了 eVLP 在人体细胞中的生产性能或转导效率。通过结合有益的囊体变异,我们开发出了第五代(v5)eVLP,与之前的最佳 v4 eVLP 相比,它在培养哺乳动物细胞中的传递效力提高了 2-4 倍。对 v5 eVLP 的分析表明,这些荚膜突变优化了所需核糖核蛋白载体而非原生病毒基因组的包装和递送,并大大改变了 eVLP 的荚膜结构。这些发现表明,条形码 eVLP 的进化有可能支持改良 eVLP 的开发。
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来源期刊
Nature biotechnology
Nature biotechnology 工程技术-生物工程与应用微生物
CiteScore
63.00
自引率
1.70%
发文量
382
审稿时长
3 months
期刊介绍: Nature Biotechnology is a monthly journal that focuses on the science and business of biotechnology. It covers a wide range of topics including technology/methodology advancements in the biological, biomedical, agricultural, and environmental sciences. The journal also explores the commercial, political, ethical, legal, and societal aspects of this research. The journal serves researchers by providing peer-reviewed research papers in the field of biotechnology. It also serves the business community by delivering news about research developments. This approach ensures that both the scientific and business communities are well-informed and able to stay up-to-date on the latest advancements and opportunities in the field. Some key areas of interest in which the journal actively seeks research papers include molecular engineering of nucleic acids and proteins, molecular therapy, large-scale biology, computational biology, regenerative medicine, imaging technology, analytical biotechnology, applied immunology, food and agricultural biotechnology, and environmental biotechnology. In summary, Nature Biotechnology is a comprehensive journal that covers both the scientific and business aspects of biotechnology. It strives to provide researchers with valuable research papers and news while also delivering important scientific advancements to the business community.
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