Endogenous GFP-tagging of pmm2 does not interfere with Pmm2 function in medaka. (A) Schematic representation of the medaka pmm2 gene locus. Endogenous GFP knock-in outline at the C-terminus of the pmm2 coding sequence via CRISPR/Cas9 (scissors) and biotinylated (red stop signs) donor template [coding sequence, red boxes; untranslated region (UTR), white boxes]. Single copy integration by homology directed repair (HDR) with the right homology flank (RH) and a non-homologous end-joining (NHEJ) event of the left homology flank (LH). Transcript analysis revealed correctly spliced pmm2-GFP mRNA. (B) Representative hatchling homozygous for pmm2-GFP shows ubiquitous GFP expression and proper development.

@kpakari.bsky.social & co recapitulate the activity levels of hypomorphic mutations associated with human PMM2-congenital disorders of glycosylation in medaka. 🐟
doi.org/10.1242/bio....

#Degron #EndogenousGFPtagging #CRISPR