Warner Jacob F, Besemer Ryan, Schickle Alicia, Borbee Erin, Changsut Isabella V, Sharp Koty, Babonis Leslie S
Department of Biology and Marine Biology, UNC Wilmington, Wilmington, NC, 28409.
Feinstein School of Social and Natural Sciences, Roger Williams University, Bristol, RI 02871.
bioRxiv. 2023 Nov 16:2023.11.16.567385. doi: 10.1101/2023.11.16.567385.
Cnidarians have become valuable models for understanding many aspects of developmental biology including the evolution of body plan diversity, novel cell type specification, and regeneration. Most of our understanding of gene function during early development in cnidarians comes from a small number of experimental systems including the sea anemone, Few molecular tools have been developed for use in hard corals, limiting our understanding of this diverse and ecologically important clade. Here, we report the development of a suite of tools for manipulating and analyzing gene expression during early development in the northern star coral, We present methods for gene knockdown using short hairpin RNAs, gene overexpression using exogenous mRNAs, and endogenous gene tagging using CRISPR-mediated gene knock-in. Combined with our ability to control spawning in the laboratory, these tools make a tractable experimental system for investigative studies of coral development. Further application of these tools will enable functional analyses of embryonic patterning and morphogenesis across Anthozoa and open new frontiers in coral biology research.
刺胞动物已成为理解发育生物学诸多方面的重要模型，包括身体结构多样性的演化、新型细胞类型的特化以及再生。我们对刺胞动物早期发育过程中基因功能的了解大多来自少数实验系统，如珊瑚虫。针对硬珊瑚开发的分子工具很少，这限制了我们对这个多样且具有重要生态意义的类群的理解。在此，我们报告了一套用于操纵和分析北极星珊瑚早期发育过程中基因表达的工具的开发情况。我们介绍了使用短发夹RNA进行基因敲低、使用外源mRNA进行基因过表达以及使用CRISPR介导的基因敲入进行内源基因标记的方法。结合我们在实验室控制产卵的能力，这些工具为珊瑚发育的研究提供了一个易于处理的实验系统。这些工具的进一步应用将能够对整个珊瑚纲的胚胎模式形成和形态发生进行功能分析，并为珊瑚生物学研究开辟新的前沿领域。
