2025/09/10 by Milan Gerovac, Leandro Buhlmann, Yan Zhu +6 · 13 citations
Environmental Science · Biochemistry, Genetics and Molecular Biology · Medicine · #Bacteriophages and microbial interactions #Advanced biosensing and bioanalysis techniques #Monoclonal and Polyclonal Antibodies Research #Functional genomics #Phage display #Gene #Gene knockdown #Gene silencing #Phagemid #RNA #Genome #Bacteriophage
paper · pdf · doi:10.1038/s41586-025-09499-6
published in Nature 646(8087), 1195-1203 (Nature Portfolio)
openalex publication_date 2025/09/10 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/25
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.