Many thanks to the team!! 🤓🤓🤡🥳 @sashafanari.bsky.social @wanunupore.bsky.social

5/
So are immortalized cells “damaged”? Not quite.
They’re not perfect stand-ins, but they’re surprisingly consistent with primary cells.
We conclude that Jurkat cells are a solid model for studying ψ — just watch out for that 13%.
📖 tinyurl.com/TCellPsiRNA

4/
The twist? It’s not due to differences in pseudouridine synthase expression — enzyme levels were similar.
So what's driving site selection? Likely trans-regulatory factors or RNA structure, not just enzyme abundance.

3/ But that other 13%?
☑️ Jurkat-specific ψ-sites hit oncogenic and immune activation genes
☑️ Primary T cell–specific ψ-sites appear on trafficking and calcium signaling genes
🧬 Immortalized cells also showed more clustered ψ patterns, hinting at altered control compared to primary T cells.

2/ We were genuinely surprised by this result — we expected transformation to cause widespread disruption. But 87% of ψ-sites were shared between Jurkat and primary T cells!
Most differences came from gene expression, not ψ-site selection.
This process seems to be tightly conserved.

6/6 🙌 Future work will determine whether static sites are critical for cellular function while plastic sites fine-tune gene expression in response to environmental stressors.
Huge thanks to the team for all their hard work! @wanunupore.bsky.social @genometdcc.bsky.social

5/6 ⚙️ Enzyme insights: TRUB1 and PUS7 levels shifted with differentiation, and KD experiments reveal unexpected coregulation—knocking down one lowers Ψ at the other’s targets. This hints at a coordinated network fine-tuning mRNA Ψ for neuronal homeostasis and stress resilience.

4/6 📊 Plasticity vs stability: ~30% of Ψ sites changed occupancy across conditions (“plastic”), while the rest remained constant (“static”). Among the plastic sites was a key Ψ site in YTHDF1—an m6A reader involved in translation, which changed in response to cellular and environmental cues.

3/6 📊 Key finding #1: Pb²⁺ treated cells had more Ψ sites transcriptome-wide but lower relative occupancy per site vs untreated/differentiated cells—suggesting a broad but shallow protective pseudouridylation response to stress.

2/6 🧠Link here: authors.elsevier.com/a/1koBC8YyDf...
Method: SH SY5Y cells were untreated, differentiated with retinoic acid, or exposed to Pb²⁺. Nanopore DRS determined site-specific, relative Ψ “occupancy”; orthogonal knockdowns (TRUB1, PUS7) and biochemical assays validated sites.

I sat on a study section that met this past Friday (2/14)