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Splice-modulating antisense oligonucleotides (ASOs) are precision RNA-based drugs that are becoming an established modality to treat human disease. Previously, we reported the discovery of ASOs that target a novel, putative intronic RNA structure to rescue splicing of multiple pathogenic variants of F8 exon 16 that cause hemophilia A. However, the conventional approach to discovering splice-modulating ASOs is both laborious and expensive. Here, we describe a novel approach that integrates data-driven RNA structure prediction and community science to discover splice-modulating ASOs. Using a splicing-deficient pathogenic variant of F8 exon 16 as a model, we show that 25% of the top-scoring molecules designed in the Eterna OpenASO challenge have a statistically significant impact on enhancing exon 16 splicing. Additionally, we show that a distinct combination of ASOs designed by Eterna players can additively enhance the inclusion of the splicing-deficient exon 16 variant. Together, our data suggests that crowdsourcing designs from a community of citizen scientists may accelerate and complement traditional avenues for the discovery of splice-modulating ASOs with potential to treat human disease. 

The Ross Sea I glaciation, marked by the northward advance of the Ross Ice Sheet (RIS) in the Ross Sea, east Antarctica, corresponds with the last major expansion of the West Antarctic Ice Sheet during the last glacial period. During its advance, the RIS was grounded along the southern Victoria Land coast, completely blocking the mouths of several of the McMurdo Dry Valleys (MDVs). Several authors have proposed that very large paleolakes, proglacial to the RIS, existed in many of the MDVs. Studies of these large paleolakes have been key in the interpretation of the regional landscape, climate, hydrology, and glacier and ice sheet movements. By far the most studied of these large paleolakes is Glacial Lake Washburn (GLW) in Taylor Valley. Here, we present a comprehensive review of literature related to GLW, focusing on the waters supplying the paleolake, signatures of the paleolake itself, and signatures of past glacial movements that controlled the spatial extent of GLW. We find that while a valley-wide proglacial lake likely did exist in Taylor Valley during the early stages of the Ross Sea I glaciation, during later stages two isolated lakes occupied the eastern and western sections of the valley, confined by an expansion of local alpine glaciers. Lake levels above ~140 m asl were confined to western Taylor Valley, and major lake level changes were likely driven by RIS movements, with climate variables playing a more minor role. These results may have major implications for our understanding of the MDVs and the RIS during the Ross Sea I glaciation. 

Abstract Aquatic ecosystems - lakes, ponds and streams - are hotspots of biodiversity in the cold and arid environment of Continental Antarctica. Environmental change is expected to increasingly alter Antarctic aquatic ecosystems and modify the physical characteristics and interactions within the habitats that they support. Here, we describe physical and biological features of the peripheral ‘moat’ of a closed-basin Antarctic lake. These moats mediate connectivity amongst streams, lake and soils. We highlight the cyclical moat transition from a frozen winter state to an active open-water summer system, through refreeze as winter returns. Summer melting begins at the lakebed, initially creating an ice-constrained lens of liquid water in November, which swiftly progresses upwards, creating open water in December. Conversely, freezing progresses slowly from the water surface downwards, with water at 1 m bottom depth remaining liquid until May. Moats support productive, diverse benthic communities that are taxonomically distinct from those under the adjacent permanent lake ice. We show how ion ratios suggest that summer exchange occurs amongst moats, streams, soils and sub-ice lake water, perhaps facilitated by within-moat density-driven convection. Moats occupy a small but dynamic area of lake habitat, are disproportionately affected by recent lake-level rises and may thus be particularly vulnerable to hydrological change. 

Abstract Pathogenic variants in the human Factor VIII (F8) gene cause Hemophilia A (HA). Here, we investigated the impact of 97 HA-causing single-nucleotide variants on the splicing of 11 exons from F8. For the majority of F8 exons, splicing was insensitive to the presence of HA-causing variants. However, splicing of several exons, including exon-16, was impacted by variants predicted to alter exonic splicing regulatory sequences. Using exon-16 as a model, we investigated the structure–function relationship of HA-causing variants on splicing. Intriguingly, RNA chemical probing analyses revealed a three-way junction structure at the 3′-end of intron-15 (TWJ-3–15) capable of sequestering the polypyrimidine tract. We discovered antisense oligonucleotides (ASOs) targeting TWJ-3–15 partially rescue splicing-deficient exon-16 variants by increasing accessibility of the polypyrimidine tract. The apical stem loop region of TWJ-3–15 also contains two hnRNPA1-dependent intronic splicing silencers (ISSs). ASOs blocking these ISSs also partially rescued splicing. When used in combination, ASOs targeting both the ISSs and the region sequestering the polypyrimidine tract, fully rescue pre-mRNA splicing of multiple HA-linked variants of exon-16. Together, our data reveal a putative RNA structure that sensitizes F8 exon-16 to aberrant splicing.