Introduction In Xenopus laevis oocytes an absence of transcription occurs in early development from before the resumption of meiosis in stage VI oocytes, through the maternal-zygotic transition (MZT) at almost 4000 cells, requiring new protein synthesis to be directed by stored mRNAs. Several modifications control the translation of these mRNAs including cytoplasmic polyadenylation and 2’-O methylation1. Mechanics of cytoplasmic polyadenylation are well established, but 2’-O methylation involvement is less clear. We are interested in the mechanism through which this modification might act in regulation of translation and are investigating candidate proteins that recognize the presence or absence of this molecular mark. The interferon induced with helicase C domain 1 (Ifih1 or Mda5) protein is known to bind RNAs based on their 2’-O methylation state2. The ifih1 transcript has been identified in the stage VI oocyte, implying possible involvement in oocyte maturation, and suggesting that the Ifih1 protein could be involved in translational regulation. An alternative function may be in developmental immunology as Ifih1 is typically responsible for recognition of dsRNA and the activation of cytokines. Little is known regarding immune responses during early Xenopus embryogenesis, but as Ifih1 is known to target (+) ssRNA viruses, such as coronaviruses, determining the role of Ifih1 in Xenopus laevis development may help better understand the protein’s overall function. Discussion While research was halted due to the University shutdown, we have indication that we have cloned the ifih1 mRNA, and have the PIV5-V ready for transcription and injection. The following considerations are guiding our experimental design: • Ifih1 usually acts through the Mavs protein to activate interferon and cytokine transcription in response to viral infection3. If it is acting as an immune sentinel it will activate through a novel mechanism as there is no transcription at this stage • The other RLR do not seem to be expressed in early development suggesting a different developmental role for this protein • There is precedence for helicases such as Ifih1 to act as regulators of translation7 • Isolation of tagged Ifih1 and binding partners, (protein and mRNA) should establish the pathway that the protein is acting in during development Elizabeth C. Devaney, Serena Iacovelli, Morgan Nelson, & Daron C. Barnard Department of Biology, Worcester State University Functional Characterization of Ifih1 and Dhx29 During Early Xenopus laevis Development Literature cited 1. Kuge, H., & Richter, J. D. (1995). Cytoplasmic 3“ poly(A) addition induces 5” cap ribose methylation: implications for translational control of maternal mRNA. The EMBO Journal, 14(24), 6301. 2. Züst, R., et al. (2011). Ribose 2′-O-methylation provides a molecular signature for the distinction of self and non-self mRNA dependent on the RNA sensor Mda5. Nature Immunology, 12(2), 137–143. 3. Dixit, E., and Kagan, J. C. (2013). Intracellular Pathogen Detection by RIG-I- Like Receptors. Adv Immunol. 117, 99–125 4. Xenbase (http://www.xenbase.org/, RRID:SCR_003280) 5. Session, A.M., et al. (2016). Genome evolution in the allotetraploid frog Xenopus laevis. Nature 538, 336-343. 6. Ramachandran, A. and Horvat, C.M. (2010). Dissociation of Paramyxovirus Interferon Evasion Activities: Universal and Virus-Specific Requirements for Conserved V Protein. J. Virol., 84 (21): 11152-11163. 7. Bourgeois, C. F., Mortreux, F., and Auboeuf, D. (2016). The multiple functions of RNA helicases as drivers and regulators of gene expression. Nat Rev Mol Cell Biol, 17, 426-438. Acknowledgments The authors would like to thank Worcester State University for the Faculty Scholarship and Creativity grant awarded to DCB to fund the research, the Office of University Advancement for the Student Research, Scholarship & Creativity Activity grant awarded to ECD to attend the 2020 TAGC, and Xenbase for the available Xenopus resources. O