Get ready to dance, our paper – Phage DisCo: targeted discovery of bacteriophages by co-culture – has been pre-printed! 🪩 This has been a really fun project to work on with @sianowen.bsky.social, @baym.lol, @nquinoneso.bsky.social, and our two talented undergrads Kesther and Carmen! - ThreadSky

ellie-rand.bsky.social • 92 days ago

Get ready to dance, our paper – Phage DisCo: targeted discovery of bacteriophages by co-culture – has been pre-printed! 🪩 This has been a really fun project to work on with @sianowen.bsky.social, @baym.lol, @nquinoneso.bsky.social, and our two talented undergrads Kesther and Carmen!

Reposted from bioRxiv Microbiology

Phage DisCo: targeted discovery of bacteriophages by co-culture https://www.biorxiv.org/content/10.1101/2024.11.22.624878v1

Comments

phagehunterkw.bsky.social•92 days ago

Congratulations to all! Great work!🎉🎉🎉 (still reading!)

baym.lol•92 days ago

🦠🧪

paulbriley.bsky.social•92 days ago

Why don't you follow Russian or Georgian methods of isolating bacteriophages? They've been doing this type of work for decades already.

baym.lol•92 days ago

Interesting, do you have a link or reference you could share to a similar technique? I’d love to see it

ellie-rand.bsky.social•92 days ago

While it is widely recognized that phage diversity is a valuable resource, techniques to sample phages have not changed in almost 100 years. These traditional methods tend to oversample abundant phages and miss rare phages where most of the diversity is harbored. This adds a lot of work! 2/

ellie-rand.bsky.social•92 days ago

What if, instead of sampling hundreds of phages, we could just make the right ones glow? Enter Phage Discovery by Co-culture (Phage DisCo)! Simply, this method combines two strains: a wildtype and a modified strain, each fluorescently tagged. Now, fluorescent signal will mark plaques of interest. 3/

ellie-rand.bsky.social•92 days ago

Phage DisCo can be further multiplexed with three bacterial strains in co-culture, and, when tested with characterized phages, the fluorescent signal shined! Maybe you can see where the disco ball comparison comes in here. 🪩 4/

ellie-rand.bsky.social•92 days ago

What if we dug into a sample a little more “wild” and a little less characterized? We combined our strains with a wastewater sample (thank you citizens of Boston for your ~contributions~), and again we found fluorescent hits! Each was isolated and characteristics confirmed on monoculture lawns. 5/

ellie-rand.bsky.social•92 days ago

What about other bacterial factors that phages interact with during infection? To test this, we co-cultured a wildtype E. coli with an isogenic strain that had GmrSD, a modification-dependent restriction enzyme, introduced. If GmrSD provides defense against a phage, we expect that strain to grow. 6/

ellie-rand.bsky.social•92 days ago

Yet again, the DisCo ball (fluorescence) led us quickly to phages of interest. The hits were isolated and confirmed by plaque assays on monoculture lawns of each strain. 7/

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