Capped off the trip with a visit to the lovely group at the Kampmann lab @kampmann.bsky.social in San Francisco full of awesome discussions about all their cool screens and presenting some of our data too.

Waiting for flight back to Oz and reflecting on the awesome @keystonesymposia.bsky.social last week on #RNA mediated regulation of #immunity. Here are a few highlights with links to the published or preprinted papers which you should definitely check out if you haven't yet ...

#RNAsky #Immunosky

The prevailing model is that a set of A-to-I edited RNAs become immunogenic when editing is reduced, and that collectively these increase the dsRNA load to pass the threshold for MDA5 activation. Our model is that in the absence of GGNBP2 the threshold is not reached and thus suppresses the pathway.

.... when we looked at A-to-I edited RNAs, we found a global decrease in their expression in the cytoplasm of GGNBP2 KO cells. We propose that GGNBP2 changes the balance of dsRNA between the nucleus and cytoplasm to control MDA5 activation. 10/X

So, we did RNAseq on total and cytoplasmic/nuclear RNA to see what was going on. Consistent with the qPCR – we saw virtually no gene expression changes in the GGNBP2 KO following loss of editing and a complete block in ISG induction. 8/X

Our top hits were GGNBP2, CNOT10 and CNOT11. The three proteins interact and are top co-dependencies in Depmap suggesting a functional link across cell types.

Incredibly, KO of GGNBP2 rescues editing-deficient cells even better than KO of MDA5, and completely blocked induction of ISGs by qPCR! 6/X

We performed a genome-wide CRISPR KO screen to look for outgrowth of KO alleles that would rescue editing deficiency.

Surprisingly, 3 genes enriched far higher than all others and had not been previously linked to dsRNA/IFN pathways before! 5/X