Eye phenotypes were dramatic: embryos developed two, one, or no eyes. Disruption of the pax6 homeodomain inhibitted eye formation, confirming its highly conserved developmental role. #LookMaNoEyes

Genotyping revealed mosaic F0 mutants. Multiple alleles, including in-frame and out-of-frame mutations, were present at the sgRNA target site, while controls remained unmodified.

Single-cell embryos were microinjected with CRISPR reagents. Embryos were held in a 3D-printed agarose dish, and dextran dye confirmed injections.

We identified a single pax6 transcript in the transcriptome, a master regulator of eye development. Alignments with other gastropod genomes revealed putative exon-intron structure, allowing exon-specific primer and sgRNA designed to target the homeodomain.

In C. fornicata larval eyes start as simple pigmented cups from ectodermal invagination, gaining pigment and a tiny lens by the veliger stage. Adults have lenticular eyes with a cornea, acellular lens, dense pigment, and layered photoreceptors.

CRISPR in molluscs is challenging: delivery is hard, embryo husbandry is hard, genomes are huge and complex. We tested whether efficient F0 knockouts could be generated in the slipper snail, Crepidula fornicata, using only a transcriptome to guide us. @vs-marine.bsky.social

Happy holidays to one and all ❄️! On the Friday before Christmas 🎄 I’m excited to share my first first-author pre-print. Are you interested in molluscs 🐚, developmental biology, CRISPR F0 KOs, Pax6, eye 👁️ development and evolution? Then here’s a thread for you. www.biorxiv.org/content/10.1...

PhDone ✅🐚✨

Shucked stack of snails (say that five times fast)

Exposed for my frantic 6 hour day of microinjecting crepidula embryos at @mblscience.bsky.social with @vs-marine.bsky.social. Heavy amounts of Ed Sheeran in the lab that day (Unofficial Sleight Lab Work music)

Looking at this note my PI left on a waste bottle and thinking--what could it mean?

I'll be at the #MalacologicalSocietyofLondon meeting this Wednesday! If anyone is interested CRISPR/cas9, snails, and cool microscopy, images, come listen to my talk and have a chat with me!

Hello all! Joining Blue sky to engage with the scientific community and to talk about #snails follow for updates on my research and science-related shenanigans!