维尔茨堡大学Jrg Vogel研究团队报道了用于噬菌体功能基因组学的可编程反义寡聚物。2025年9月10日出版的《自然》杂志发表了这项成果。

为了克服这一限制，课题组研究人员引入了一种非遗传mRNA靶向方法，将可编程反义寡聚物外源性递送，以沉默DNA和RNA噬菌体的基因。通过系统敲除巨噬体ΦKZ核心和辅助基因，结合RNA测序和显微镜分析，揭示了以前未被识别的蛋白质，这些蛋白质对噬菌体繁殖至关重要，并且在沉默后，在噬菌体和宿主反应水平上引发不同的表型。其中一个因素是RNase h样蛋白ΦKZ155（也称为Nlp2），它在感染过程中起着重要的决定作用，将保护性噬菌体核的形成与噬菌体基因组扩增联系起来。这种基于非遗传反义寡聚体的基因沉默方法有望成为噬菌体生物学分子发现的多功能工具，将有助于阐明非模型噬菌体-宿主对的防御和抗防御机制，并为优化噬菌体治疗和生物技术程序提供潜力。

据介绍，噬菌体是地球上最丰富的实体，具有巨大的遗传和表型多样性。利用这一很大程度上未开发的分子空间需要鉴定和功能表征在噬菌体-宿主界面上起作用的基因。到目前为止，这仅限于少数可以进行基因操作的噬菌体-宿主系统模型。

附：英文原文

Title: Programmable antisense oligomers for phage functional genomics

Author: Gerovac, Milan, Buhlmann, Leandro, Zhu, Yan, urica-Miti, Svetlana, Rech, Valentin, Carien, Samuel, Grfenhan, Tom, Popella, Linda, Vogel, Jrg

Issue&Volume: 2025-09-10

Abstract: Bacteriophages are the most abundant entities on earth and exhibit vast genetic and phenotypic diversity. Exploitation of this largely unexplored molecular space requires identification and functional characterization of genes that act at the phage–host interface. So far, this has been restricted to few model phage–host systems that are amenable to genetic manipulation. Here, to overcome this limitation, we introduce a non-genetic mRNA targeting approach using exogenous delivery of programmable antisense oligomers to silence genes of DNA and RNA phages. A systematic knockdown screen of core and accessory genes of the nucleus-forming jumbo phage ΦKZ, coupled to RNA-sequencing and microscopy analyses, reveals previously unrecognized proteins that are essential for phage propagation and that, upon silencing, elicit distinct phenotypes at the level of the phage and host response. One of these factors is the RNase H-like protein ΦKZ155 (also known as Nlp2), which acts at a major decision point during infection, linking the formation of the protective phage nucleus to phage genome amplification. This non-genetic antisense oligomer-based gene silencing method promises to be a versatile tool for molecular discovery in phage biology, will help to elucidate defence and anti-defence mechanisms in non-model phage–host pairs, and offers potential for optimizing phage therapy and biotechnological procedures.

DOI: 10.1038/s41586-025-09499-6

Source: https://www.nature.com/articles/s41586-025-09499-6