Search

Expression of exon-specific isoforms from alternatively spliced mRNA is a fundamental mechanism that substantially expands the proteome of a cell. However, conventional methods to assess alternative splicing are either consumptive and work-intensive or do not quantify isoform expression longitudinally at the protein level. Here, we therefore developed an exon-specific isoform expression reporter system (EXSISERS), which non-invasively reports the translation of exon-containing isoforms of endogenous genes by scarlessly excising reporter proteins from the nascent polypeptide chain through highly efficient, intein-mediated protein splicing. We applied EXSISERS to quantify the inclusion of the disease-associated exon 10 in microtubule-associated protein tau (MAPT) in patient-derived induced pluripotent stem cells and screened Cas13-based RNA-targeting effectors for isoform specificity. We also coupled cell survival to the inclusion of exon 18b of FOXP1, which is involved in maintaining pluripotency of embryonic stem cells, and confirmed that MBNL1 is a dominant factor for exon 18b exclusion. EXSISERS enables non-disruptive and multimodal monitoring of exon-specific isoform expression with high sensitivity and cellular resolution, and empowers high-throughput screening of exon-specific therapeutic interventions., Truong et al. developed a cell-based reporter system, EXSISERS, that enables non-invasive quantification of the protein expression levels of exon-specific isoforms via intein-mediated protein splicing.

The Caribbean island of Jamaica is home to a lineage of crabs (Crustacea: Decapoda: Brachyura: Sesarmidae) which have colonized the most unusual terrestrial habitats. Today, they can be found nesting in bromeliad leaf axils or empty snail shells, scurrying on the forest floor or among dry rock rubble, climbing on walls of deep cave systems, or digging burrows in the banks of mountain streams. In order to facilitate survival of their offspring in these remarkable and new habitats, complex behavioral adaptations have evolved, including feeding and protecting the offspring, and transport of empty snail shells into the nursery to buffer the pH and provide the necessary calcium carbonate [summarized in Diesel (Proc Roy Soc Lond B 264:1403–1406, 1997), Diesel and Schubart (Evolutionary ecology of social and sexual systems: crustaceans as model organisms. Oxford University Press, New York, pp 365–386, 2007)]. No other American sesarmid crab species shows this degree of terrestriality and elaborate brood care behavior. Instead, the most related species typically are inhabitants of marshes and mangroves and reproduce by releasing their larvae into the ocean, without any additional paternal obligation. Molecular phylogenetic analyses revealed that the Jamaican endemic sesarmids are monophyletic and thus the outcome of a single colonization event, which took place approximately 4.5 million years ago and was followed by a rapid radiation, speciation, and adaptation to ecological niches, which have not been used by crabs in any other parts of the world and required evolution of complex behavioral strategies to render survival in these habitats possible (Schubart et al. Nature 393:363–365, 1998a). This system can be considered a classic example of an adaptive radiation of comparatively young age. It calls for research concerning the mechanisms and the pace of evolution to generate these highly specialized forms. In the present paper, we provide new data on evolutionary processes at the intraspecific level, which may help to understand the potential of diversification in these crabs. Furthermore, published and unpublished data available for these specialized crabs are summarized, giving evidence for the astounding diversity of evolutionary strategies and the dimension of biodiversity beyond the species level.

Due to their long-term ecological stability, ancient lakes represent very interesting systems for studying differentiation and speciation processes. High degrees of endemism and specialisation are characteristic features of their fauna. The Malili lakes from the Indonesian island Sulawesi are considered an ancient limnic system, and recent research has increased the number of recognized species and the knowledge about ecological and morphological diversification within a variety of animal taxa. Here we show that the local endemic gecarcinucid freshwater crabs are more differentiated than qualitative morphological characters and the current taxonomy may indicate. The morphologically and ecologically well characterised species Nautilothelphusa zimmeri Balss, 1933 consists of two allopatric populations with one of the groups being more closely related to sympatric Parathelphusa ferruginea Chia and Ng, 2006 according to mitochondrial genetic data. One haplotype is even shared by both species. Nevertheless, in this study lack of gene flow between both species is demonstrated. To our knowledge, this is the first report of DNA sequence identity between two different genera along the same cytochrome oxidase I gene segment that is being proposed for barcoding studies.