Thank you Zamin!

Thanks Ellinor, I'm happy to have my CRISPR child released out into the world!

Huge thank you to the lovely co-authors who made this work possible @taylorlabgroup.bsky.social Edze Westra @bridgetwatson.bsky.social @pinkpetri.bsky.social @gretelwaugh.bsky.social and Yueyi Cai

This nutrient dependent efficacy of CRISPR-Cas interference on phage was a really cool finding and potentially opens up bigger questions about what we might be missing when we screen phage and defence systems in only optimal growth conditions.

We ran all our initial tests in optimal high nutrient conditions. However, when we re-tested some of our CRISPR-impervious phage again in low nutrient conditions we found that two of these phages could no longer resist CRISPR defences, allowing the bacteria to grow unaffected by phage lysis!

Interestingly we found that CRISPR worked great against a variety of temperate and virulent phages across a range of infection metrics. However, a few phages were resistant to CRISPR, some which made sense (jumbo phage) and others not.

Once integrated into the P. aeruginosa genome the synthetic CRISPR-Cas system worked great against plasmids, so we turned our system against a panel of phage to see if, when given targeting spacers, we could find patterns as to what phages CRISPR does and doesn’t work against.

Because I put restriction sites around all parts of the CRISPR-Cas genes, including the spacers, it made it simple to swap out the spacers with cheap primers before integrating the whole system back into the genome of P. aeruginosa, thus allowing us to target this system against any phage we wanted.

We were interested into diving deeper on understanding how useful CRISPR-Cas systems are to the bacteria that carry or acquire them. So at the start of my PhD I spent many hours creating a synthetic, minimal, modular version of the type IF CRISPR-Cas system of Pseudomonas aeruginosa.

Such a cool story! So exciting to see it out after seeing your talk on it ☺️

Best of luck with your next adventure!!

Thanks Alice! ☺️☺️☺️