Have you ever wanted to *see* receptor activity in embryos? If so, our new preprint is for you! In it, we showcase a new live-cell biosensor for visualizing receptor tyrosine kinase activity in living embryos – pYtags! www.biorxiv.org/content/10.1... - ThreadSky

emkolen.bsky.social • 51 days ago

Have you ever wanted to *see* receptor activity in embryos? If so, our new preprint is for you! In it, we showcase a new live-cell biosensor for visualizing receptor tyrosine kinase activity in living embryos – pYtags!

https://www.biorxiv.org/content/10.1101/2025.01.06.631605v1

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lumasullo.bsky.social•32 days ago

That’s super cool work!! Congratulations! 🎉

stearnslab.bsky.social•49 days ago

Congratulations on the beautiful work, Emily!

emkolen.bsky.social•49 days ago

Thanks Tim!!

emkolen.bsky.social•51 days ago

There are lots of reasons to want to visualize receptor activity in vivo – measuring morphogen gradients, detecting subcellular receptor activation during chemotaxis, understanding signaling dynamics (i.e. ERK waves), etc etc. But we need new tools!

emkolen.bsky.social•51 days ago

The Toettcher lab recently developed a new class of biosensors for receptor tyrosine kinases (RTKs) called pYtags. They work beautifully in mammalian cell culture. But could pYtags be used to measure endogenous RTK activity in embryos?

https://elifesciences.org/articles/82863

emkolen.bsky.social•51 days ago

pYtags use an exceptionally specific and orthogonal pTyr / SH2 binding pair (ITAM + ZtSH2) to recruit a fluorescently tagged protein to phosphorylated RTKs.

cellmorphosero.bsky.social•50 days ago

Would it be possible to tag non-receptor TKs (that are membrane localised)?

emkolen.bsky.social•50 days ago

Possibly! If the phosphorylated substrate was membrane localized and could be tagged with the ITAM. Jak/Stat receptors would be a good example

emkolen.bsky.social•51 days ago

We were particularly interested in Torso, the RTK controlling terminal ERK signaling at the poles of the fly embryo. I tagged Torso with the pYtag and saw gorgeous signal at the poles just where we expected it! pYtags work in embryos!

emkolen.bsky.social•51 days ago

And pYtags didn’t just work for Torso! We also made pYtags for EGFR and FGFR/Btl, and those worked just as well. The pYtag strategy is so powerful because it is generalizable to any RTK of interest.

emkolen.bsky.social•51 days ago

We then asked what we could learn about Torso by comparing Torso activity with downstream ERK activity. Two things surprised us, highlighting that we can’t always accurately infer receptor activity from downstream signaling.

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