I-Scei Meganuclease-mediated transgenesis in the acorn worm, Saccoglossus kowalevskii
I-Scei Meganuclease-mediated transgenesis in the acorn worm, Saccoglossus kowalevskii
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DOI：
10.1016/j.ydbio.2018.10.022
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发表时间：
2019-01-01
影响因子：
2.7
通讯作者：
Lowe, Christopher J.
中科院分区：
文献类型：
--
作者：
Minor, Paul J.;Clarke, D. Nathaniel;Lowe, Christopher J.
半脊索动物是一门海洋无脊椎动物后口动物，与脊索动物密切相关，是最有前途的模型之一，以提供见解早期后口动物的进化。半脊索动物的基因组，Saccoglossus kowalevskii，揭示了一个广泛的非编码元件保守的所有三个后口门。这些假定的调控元件的功能表征和跨门比较将使增强子进化的更好的理解，以及随后如何在基因调控的变化引起形态创新。在这里，我们描述了一种有效的方法转基因的非编码元件的特征在S。科瓦列夫斯基。我们首先测试的能力的I-SceI转基因系统驱动无处不在或区域化的基因表达，并标记特定的细胞类型。最后，我们确定了一个最小的启动子，可以用来测试在S。kowaleuskii。这项工作表明，这种I-SceI转基因技术，再加上对染色质可及性的理解，可以成为一个强大的工具，用于研究如何在基因调控机制的进化变化，有助于多样化的身体计划在后口动物。
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. kowaleuskii. 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.