Approaches for bacteriophage genome engineering

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

doi:10.1016/j.tibtech.2022.08.008

Approaches for bacteriophage genome engineering

Trends Biotechnol. 2023 May;41(5):669-685. doi: 10.1016/j.tibtech.2022.08.008. Epub 2022 Sep 15.

Authors

Marina Mahler¹, Ana Rita Costa², Sam P B van Beljouw², Peter C Fineran³, Stan J J Brouns⁴

Affiliations

¹ Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand; Department of Bionanoscience, Delft University of Technology, Delft, the Netherlands.
² Department of Bionanoscience, Delft University of Technology, Delft, the Netherlands; Kavli Institute of Nanoscience, Delft, the Netherlands.
³ Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand; Bioprotection Aotearoa, University of Otago, Dunedin, New Zealand; Genetics Otago, University of Otago, Dunedin, New Zealand.
⁴ Department of Bionanoscience, Delft University of Technology, Delft, the Netherlands; Kavli Institute of Nanoscience, Delft, the Netherlands. Electronic address: [email protected].

PMID: 36117025
DOI: 10.1016/j.tibtech.2022.08.008

Abstract

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.

Keywords: CRISPR-Cas; homologous recombination; phage engineering; rebooting; recombineering.

Publication types

Review
Research Support, Non-U.S. Gov't

MeSH terms

Bacteria / genetics
Bacteriophages* / genetics
CRISPR-Cas Systems
Genome, Viral