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CRISPR/Cas9 Mutagenesis Reveals A Role For ABCB1 In Gut Immune Responses To Vibrio Diazotrophicus In Sea Urchin Larvae

Travis J. Fleming, Catherine S. Schrankel, Himanshu Vyas, Hannah D. Rosenblatt, Amro Hamdoun

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ABSTRACT The ABC transporter ABCB1 plays an important role in the disposition of xenobiotics. Embryos of most species express high levels of this transporter in early development as a protective mechanism, but its native substrates are not known. Here, we used larvae of the sea urchin Strongylocentrotus purpuratus to characterize the early life expression and role of Sp-ABCB1a, a homolog of ABCB1. The results indicate that while Sp-ABCB1a is initially expressed ubiquitously, it becomes enriched in the developing gut. Using optimized CRISPR/Cas9 gene editing methods to achieve high editing efficiency in the F0 generation, we generated ABCB1a crispant embryos with significantly reduced transporter efflux activity. When infected with the opportunistic pathogen Vibrio diazotrophicus, Sp-ABCB1a crispant larvae demonstrated significantly stronger gut inflammation, immunocyte migration and cytokine Sp-IL-17 induction, as compared with infected control larvae. The results suggest an ancestral function of ABCB1 in host–microbial interactions, with implications for the survival of invertebrate larvae in the marine microbial environment.