Unbiased discovery of Novel Regulators of the Cranial Neural Crest

描述（由申请人提供）：这项工作的长期目标是发展对神经嵴发育的机制性理解，神经嵴是脊椎动物进化的核心干细胞样祖细胞群体，以及了解这种细胞类型的异常发育如何导致先天性畸形和疾病（如癌症）。我们试图了解在一个机械和生物化学水平的干细胞群体是如何形成的，是什么控制其多能性的维护，这些细胞的迁移和侵入行为是如何调节的，并最终，如何个别神经嵴细胞定向采取特定的衍生命运。该提案的总体目标是确定NC-GRN内的新组件和相互作用。更具体地说，它试图使用下一代测序，识别神经嵴所需的miRNA和关键神经嵴转录因子Snail的下游靶标。我们假设miRNA在调节神经嵴发育中起重要作用，包括其产生颅面骨骼的能力。在神经嵴发育过程中表达的miRNAs的身份及其在该组织中的作用在很大程度上仍然未知。在NC-GRN的核心成分中，已知Snail家族的转录因子在其他模式生物的神经嵴发育过程中发挥着重要作用。因此，该提议还将通过产生用于以时间控制的方式消融Snail功能的遗传工具来确定作为Snail功能的结果，什么miRNAS在神经嵴细胞中表达或从神经嵴细胞排除。该工具还将用于在全基因组范围内识别神经嵴发育关键阶段的Snail转录靶点。这项工作对于了解神经嵴细胞和神经嵴衍生物的正常发育，包括颅面骨骼，以及了解广泛的神经嵴相关先天性缺陷具有重要意义。

DESCRIPTION (provided by applicant): The long-term goal of this work is to develop a mechanistic understanding of the development of the neural crest, a stem cell like progenitor population central to the evolution of the vertebrates, as well as to understand how congenital malformations and diseases such as cancer result from the aberrant development of this cell type. We seek to understand at a mechanistic and biochemical level how the stem cell population is formed, what controls the maintenance of its multipotency, how the migratory and invasive behavior of these cells is regulated, and ultimately, how individual neural crest cells are directed to adopt specific derivative fates. The overall goal of this proposal is to identify novel components and interactions within the NC-GRN. More specifically it seeks to, using next generation sequencing, identify miRNAs required for neural crest and downstream targets of the key neural crest transcription factor Snail. We hypothesize that miRNAs play an essential role in regulating neural crest development, including its ability to give rise to the craniofacial skeleton. The identity of the miRNAs expressed during neural crest development and their roles in this tissue remain largely unknown. Of the core components of the NC-GRN the Snail family of transcription factors is known to play reiterative roles during neural crest development in other model organisms. This proposal will therefore also determine what miRNAS are expressed in or excluded from neural crest cells as a consequence of Snail function by generating a genetic tool for ablating Snail function in a temporally controlled manner. This tool will also be used to identify, on a genome wide scale, Snail transcriptional targets during key stages of neural crest development. This work is of high significance for understanding the normal development of neural crest cells and neural crest derivatives, including the craniofacial skeleton, and for understanding a broad range of neural crest linked congenital defects.