Engineering Campylobacter phages using CRISPRi at Københavns Universitet

Introduction: CRISPR-Cas has been used in E. coli and other bacteria to genetically engineer bacteriophages (phages). However, such plasmid-based systems are not compatible with the major zoonotic pathogen Campylobacter jejuni. Recently, our collaborators in the UK developed a CRISPRi platform located on the bacterial chromosome of C. jejuni. However, it has not been studied if CRISPRi can be used to genetically engineer phages infecting C. jejuni.

Aim: Genetically engineer Fletchervirus phages infecting C. jejuni using CRISPRi as a proof-of-concept.

Your work: You will use CRISPRi-modified C. jejuni strains sensitive to Fletchervirus phages with guides targeting one of the receptor binding proteins (RBP2) to as a proof-of-concept show that you can modify phages using this technology. If successful, you will design and modify CRISPRi strains with new guides targeting other phage genes of interest. Furthermore, you will learn how to work with pathogenic bacteria and get GMOII lab experience as well as be working in an international research lab.

Methods: Hands-on C. jejuni and bacteriophage experiments, CRISPRi guide design and application in C. jejuni, basic molecular techniques, bioinformatic genome analysis, etc.

Promoter and tutor in the lab: Assistant professor Martine Camilla Holst Sørensen

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