Sun Dennis A, Bredeson Jessen V, Bruce Heather S, Patel Nipam H
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
Marine Biological Laboratory, Woods Hole, MA 02543, USA.
Development. 2022 Jun 1;149(11). doi: 10.1242/dev.200793. Epub 2022 Jun 10.
Emerging research organisms enable the study of biology that cannot be addressed using classical 'model' organisms. New data resources can accelerate research in such animals. Here, we present new functional genomic resources for the amphipod crustacean Parhyale hawaiensis, facilitating the exploration of gene regulatory evolution using this emerging research organism. We use Omni-ATAC-seq to identify accessible chromatin genome-wide across a broad time course of Parhyale embryonic development. This time course encompasses many major morphological events, including segmentation, body regionalization, gut morphogenesis and limb development. In addition, we use short- and long-read RNA-seq to generate an improved Parhyale genome annotation, enabling deeper classification of identified regulatory elements. We discover differential accessibility, predict nucleosome positioning, infer transcription factor binding, cluster peaks based on accessibility dynamics, classify biological functions and correlate gene expression with accessibility. Using a Minos transposase reporter system, we demonstrate the potential to identify novel regulatory elements using this approach. This work provides a platform for the identification of novel developmental regulatory elements in Parhyale, and offers a framework for performing such experiments in other emerging research organisms.
新兴的研究生物体使得利用传统“模式”生物体无法开展的生物学研究成为可能。新的数据资源能够加速对此类动物的研究。在此,我们展示了用于夏威夷长腹剑水蚤这种双甲目甲壳动物的新功能基因组资源,有助于利用这种新兴研究生物体探索基因调控进化。我们使用全基因组转座酶可及染色质测序(Omni-ATAC-seq)在夏威夷长腹剑水蚤胚胎发育的广泛时间进程中鉴定全基因组范围内的可及染色质。这个时间进程涵盖许多主要的形态发生事件,包括体节形成、身体区域化、肠道形态发生和肢体发育。此外,我们使用短读长和长读长RNA测序来改进夏威夷长腹剑水蚤的基因组注释,从而能够对已鉴定的调控元件进行更深入的分类。我们发现差异可及性、预测核小体定位、推断转录因子结合、基于可及性动态对峰进行聚类、分类生物学功能并将基因表达与可及性相关联。使用minos转座酶报告系统,我们证明了利用这种方法鉴定新型调控元件的潜力。这项工作为在夏威夷长腹剑水蚤中鉴定新型发育调控元件提供了一个平台,并为在其他新兴研究生物体中开展此类实验提供了一个框架。