Approaches for bacteriophage genome engineering

Marina Mahler, Ana Rita Costa, Sam P.B. van Beljouw, Peter C. Fineran, Stan J.J. Brouns*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

9 Citations (Scopus)
21 Downloads (Pure)


In recent years, bacteriophage research has been boosted by a rising interest in using phage therapy to treat antibiotic-resistant bacterial infections. In addition, there is a desire to use phages and their unique proteins for specific biocontrol applications and diagnostics. However, the ability to manipulate phage genomes to understand and control gene functions, or alter phage properties such as host range, has remained challenging due to a lack of universal selectable markers. Here, we discuss the state-of-the-art techniques to engineer and select desired phage genomes using advances in cell-free methodologies and clustered regularly interspaced short palindromic repeats-CRISPR associated protein (CRISPR-Cas) counter-selection approaches.

Original languageEnglish
Pages (from-to)669-685
Number of pages17
JournalTrends in Biotechnology
Issue number5
Publication statusPublished - 2023


  • CRISPR-Cas
  • homologous recombination
  • phage engineering
  • rebooting
  • recombineering


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