iiinova — where your digital self keeps evolving

Reposted by Amanda Shilton

Alt CDC (they/them) @altcdc.altgov.info · 08/02/2025

Stay strong my friends. Rest. Recharge. Dig deep. We need each other and the country needs us. I have no idea what's awaiting me, or what will happen when this all ends. For the moment I know this: there are sick people and they need curing. Albert Camus, The Plague

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

So much gratitude to Luciano and the Marraffini Lab for the unwavering support over the past few years, and to Naama @naamaaviram.bsky.social for being such an incredible colleague!! See supplemental videos for some cool infection/growth arrest displays!!

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

Finally, we investigated the evolutionary importance of PL spacers since they come at such a fitness cost. We showed that a triggered growth arrest provides broad-spectrum immunity and can protect cells from later infection by untargeted phages!

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

Without Cas10 activity, all spacers behave like PL suggesting that Cas10 normally prevents growth arrest in PE. Compared to PE, PL spacers are rarely acquired so there must be a mechanism to retain them. We wondered if Cas10 could allow eventual recovery from growth arrest.

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

Mutating the accessory Type III RNase, Csm6, showed that downstream PE spacers, in addition to PL, fail to provide immunity without Csm6 activity. Downstream PE regions become increasingly dependent on this RNase but do not seem to trigger the prolonged arrest seen with PL.

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

Growth assays and microscopy showed that targeting PE transcripts allows uninhibited growth of infected cells but targeting PL transcripts causes a growth arrest allowing only the uninfected cells to proliferate. Surprisingly, some regions of downstream PE behave like PL.

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

We characterized the immune response mediated by spacers across the entire phage genome. RNA-seq revealed that phage transcription is controlled by 2 promoters. Spacers targeting early transcripts (PE) are enriched, but those targeting late transcripts (PL) also provide immunity.

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Amanda Shilton @akshilton.bsky.social · 04/12/2024

Incredibly proud to share my thesis work, co-led by @naamaaviram.bsky.social available now at Cell Host Microbe! Cells use growth arrest as a defense against stress and viruses. Here we thoroughly characterize the arrest associated with CRISPR Type III immunity. www.cell.com/cell-host-mi...

cell.com

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