MicroRNA evolution in placental mammals: Unravelling conservation and divergence in their regulatory mechanisms in early pregnancy in different placen

背景：在真兽哺乳动物中，胎盘作为一个器官的出现与基因组中新的调节机制的出现相协调。曾经这样的例子是胎盘哺乳动物茎系中的一组特定的microRNAs （miRNAs）。mirna在子宫内膜和胎盘功能中发挥重要作用，以建立成功的妊娠。然而，目前尚不清楚这些胎盘哺乳动物特异性mirna的调节机制有多保守或多样化，以及它们是否将胎盘作为一个器官本身或胎盘哺乳动物生理的其他方面进行调节。目的：1 .确定不同物种胎盘哺乳动物特异性组织（子宫内膜、胎盘、乳腺）中新型胎盘哺乳动物特异性mirna的表达及其预测靶点。2 .在体外研究候选miRNA是否通过敲低或过表达以分化或保守的方式调节基因表达。3 .确定候选miRNA是否调节同一物种中相同或不同的过程，即乳腺或胎盘对基因表达的特异性调节。新颖性：这个前沿项目结合了子宫和胎盘生物学、miRNA调控和计算进化生物学，以解决miRNA如何促进胎盘哺乳动物的出现和胎盘多样性。及时性：该项目将重点关注14个microRNA，这些microRNA专门出现在所有胎盘哺乳动物的共同祖先中——由Forde博士和O'Connell博士共同监督的一名博士生生成的未发表的新数据，以及O'Connell博士等人最近发表的胎盘哺乳动物系统发育。实验方法：候选miRNA和靶表达将在代表不同胎盘形态的物种（包括牛、猪、猫、啮齿动物和有袋动物）的子宫内膜、胎盘和乳腺组织中进行评估。功能分析将通过体外敲除/添加miRNA模拟物到体外细胞系，并通过RNAseq和/或纳米lc MS/MS测量效果来验证。

Background:In eutherian mammals, the emergence of the placenta as an organ was coordinate with the emergence of novel regulatory mechanisms in the genome. Once such example is a specific cohort of microRNAs (miRNAs) at the stem lineage of placental mammals. miRNAs play an important role in uterine endometrial and placental function to establish successful pregnancy. It is unclear however, how conserved or diverse the regulatory mechanisms of these placental mammal-specific miRNAs are and if they regulate the placenta as an organ itself or some other aspect of placental mammal physiology.Objectives:1-To determine the expression of novel placental mammal-specific miRNAs and their predicted targets in placental mammal-specific tissues (endometrium, placenta, mammary gland) from different species. 2-To investigate if candidate miRNAs regulate gene expression in a divergent or conserved manner via knockdown or overexpression of miRNA's in in vitro. 3-To determine if candidate miRNA (s) regulate the same or different processes in the same species i.e. mammary gland or placental specific regulation of gene expression. Novelty:This cutting-edge project combines uterine and placental biology, miRNA regulation and computational evolutionary biology to address how miRNAs contributed to the emergence of placental mammals and placental diversity.Timeliness:The project will focus on 14 microRNA's that arose specifically in the common ancestor of all placental mammals - novel unpublished data generated by a PhD student co-supervised by Dr Forde and Dr O'Connell, as well as placental mammal phylogeny which has recently been published by Dr O'Connell et al. Experimental Approaches: Candidate miRNA and target expression will be assessed in endometrial, placental and mammary gland tissues from species representing distinctive placental morphologies including bovine, porcine, feline, rodent, and marsupials. Functional analysis will be validated by in vitro knock down/addition of miRNA mimics to in vitro cell lines and effects measured by RNAseq and/or nano-LC MS/MS.