It is finally out! Muntathar Al-Shimary, @doudna-lab.bsky.social , @cresslab.bsky.social and I present a gene knockdown tool that works in diverse bacteriophages: CRISPRi through antisense RNA Targeting (CRISPRi-ART)! Check it out @naturemicrobiol.bsky.social www.nature.com/articles/s41... - ThreadSky

benadler.bsky.social • 1 day ago

It is finally out!

Muntathar Al-Shimary, @doudna-lab.bsky.social , @cresslab.bsky.social and I present a gene knockdown tool that works in diverse bacteriophages: CRISPRi through antisense RNA Targeting (CRISPRi-ART)!

Check it out @naturemicrobiol.bsky.social
https://www.nature.com/articles/s41564-025-01935-7

Comments

benadler.bsky.social•1 day ago

Bacteriophages (phage) are collectively the most abundant and diverse forms of life(?) on earth. They are predators of bacteria and are locked in a genetic “arms race” with their bacterial hosts. This has led to incredible molecular innovation... and genetic diversity.
#microsky #phagesky

benadler.bsky.social•1 day ago

This diversity between phages and their hosts is more than curiosity, *it is genetic innovation*. You can trace lots of #biotechnology (CRISPR, RM, phage display, protein expression) to a simple question: "is this gene important for phage infection and when?"

benadler.bsky.social•1 day ago

Thanks to advances in synthetic and systems biology, we have been able to do this at genome-wide scale for bacterial genes through methods like gene overexpression, knockout libraries and CRISPRi, but not for phage genomes.

...but phage genomes, collectively, have been inaccessible.

benadler.bsky.social•1 day ago

Early in my postdoc, I realized that phage genomes were surprisingly susceptible to RNA-targeting, RNA degrading Cas13 (https://www.nature.com/articles/s41564-022-01258-x)

benadler.bsky.social•1 day ago

Building upon this theme
@cresslab.bsky.social myself and Muntathar teamed up to use catalytically deactivated Cas13d to targetedly knock down protein expression in phage genomes (this is CRISPRi-ART) by targeting the ribosome binding sites of phage genes.

benadler.bsky.social•1 day ago

If you target an important (or "fit") gene, you can observe both mild and dramatic phenotypes by simply changing a crRNA sequence (ie ~30bp) and works across a large diversity of phages.

benadler.bsky.social•1 day ago

We originally preprinted this technology in mid 2023 and have had a lot of updates since (will update thread later today)

Original post: https://bsky.app/profile/benadler.bsky.social/post/3k7rjmy6pvs2r