Roadmap for Phage technologies

What are we trying to achieve and why? What is the problem we are trying to solve?

Using phages as a delivery system (including to deliver i) factors that modulate AMR by targeting AMR plasmids ii) technology such as CRISPR-Cas including for modulation of virulence factors, iii) antibodies and iv) antimicrobial peptides).

What are the scientific and technological challenges (knowledge gaps needing to be addressed)?

Stability of phage genetics.
Mechanisms of function.
Plasmid stability and retransformation.
Bacterial selection and phage resistance.
Efficacy in pure culture and complex communities (i.e., microbiome).
Better engineering needed to construct mutants – A robust retargetable phage platform is needed for that.
Phages used to deliver plasmid disrupting genes may be considered GM organisms.

What approaches could/should be taken to address the research question?

High throughput systems – CRISPR-Cas technology to edit out parts of the phages would be useful.
Improvement in the application of synthetic biology to phage biology and for example exploiting Gibson assemblies and rebooting or CRISPR technology could help considerably.
Experimental design requires sufficient replication to account for variation.
Projects could include a testing pipeline of increasing complexity: pure culture > artificial or ex vivo communities > animal studies.
Experimental design requires sufficient replication to account for variation.
Cell free platform.
Might use simple phages such as small filamentous phage (e.g. M13) for delivery.

What else needs to be done before we can solve this need?

[Need to have good genetic systems.] Development of genetic systems for all phages.
[Need to genuinely compare good native phages to engineered ones.]
Mechanics of more widely applying editing systems.

Existing knowledge including successes and failures