Inactivating members of the PAG gene family by utilizing cytdine deaminase CRISPR technology
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Pregnancy-associated glycoproteins (PAGs) are aspartic proteinases produced by trophoblasts at the fetal-maternal interface. The exact functions of PAGs are unknown. This experiment aims to elucidate PAG function via the ablation of select porcine PAGs in embryos. PAG null embryos were generated via cytidine deaminase base editor. 2 guides targeting homology in 9 of the 12 PAGs were injected into zygotes via microinjection. Subsequent blastocysts were transferred to surrogates and then collected on various days of pregnancy (day 25, day 18). Day 25 yielded no pregnancies. On day 18, the conceptus was present, and PAGKO was confirmed. Histology showed aberrant vascular formation beneath the uterine epithelium. RNAseq analysis showed the down- regulation of several targets related to ECM remodeling. Porcine PAGs appear to affect proper ECM remodeling, though further research will be needed to elucidate the cellular mechanisms of these interactions.
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M.S.