I-SceI Meganuclease-mediated transgenesis in the acorn worm, Saccoglossus kowalevskii |
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Authors: | Paul J. Minor D. Nathaniel Clarke José M. Andrade López Jens H. Fritzenwanker Jessica Gray Christopher J. Lowe |
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Affiliation: | 1. Hopkins Marine Station, Department of Biology, Stanford University, 120 Oceanview Blvd., Pacific Grove, CA 93950, United States;2. Department of Systems Biology, Harvard Medical School, 200 Longwood Ave, Boston, MA 02115, United States;3. Department of Biology, Georgetown University, 411 Regents Hall, 37th and O Streets, NW, Washington DC 20057, United States |
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Abstract: | Hemichordates are a phylum of marine invertebrate deuterostomes that are closely related to chordates, and represent one of the most promising models to provide insights into early deuterostome evolution. The genome of the hemichordate, Saccoglossus kowalevskii, reveals an extensive set of non-coding elements conserved across all three deuterostome phyla. Functional characterization and cross-phyla comparisons of these putative regulatory elements will enable a better understanding of enhancer evolution, and subsequently how changes in gene regulation give rise to morphological innovation. Here, we describe an efficient method of transgenesis for the characterization of non-coding elements in S. kowalevskii. We first test the capacity of an I-SceI transgenesis system to drive ubiquitous or regionalized gene expression, and to label specific cell types. Finally, we identified a minimal promoter that can be used to test the capacity of putative enhancers in S. kowalevskii. This work demonstrates that this I-SceI transgenesis technique, when coupled with an understanding of chromatin accessibility, can be a powerful tool for studying how evolutionary changes in gene regulatory mechanisms contributed to the diversification of body plans in deuterostomes. |
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Keywords: | Hemichordate Transgenesis Enhancer Evolution |
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