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

背景：在真兽目哺乳动物中，胎盘作为一个器官的出现与基因组中新的调控机制的出现是协调的。一个这样的例子是胎盘哺乳动物干细胞谱系中的一组特定的microRNA（miRNA）。miRNAs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，目前还不清楚这些胎盘哺乳动物特异性miRNA的调控机制是如何保守或多样化的，以及它们是否调控胎盘作为一个器官本身或胎盘哺乳动物生理学的一些其他方面。目的：1-确定新的胎盘哺乳动物特异性miRNA的表达及其预测的靶点在来自不同物种的胎盘哺乳动物特异性组织（子宫内膜、胎盘、乳腺）中。2-研究候选miRNA是否通过体外敲低或过表达miRNA以发散或保守的方式调节基因表达。3-确定候选miRNA是否调节相同物种中的相同或不同过程，即基因表达的乳腺或胎盘特异性调节。新奇：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。时间：该项目将重点关注14种在所有胎盘哺乳动物的共同祖先中特异性出现的microRNA-由福尔德博士和O 'Connell博士共同监督的博士生产生的未发表的新数据，以及最近由O' Connell博士等人发表的胎盘哺乳动物遗传学。实验方法：将在来自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、胎盘和乳腺组织中评估候选miRNA和靶点表达。功能分析将通过体外敲除/向体外细胞系添加miRNA模拟物进行验证，并通过RNAseq和/或nano-LC MS/MS测量效果。

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project will focus on 14 microRNA's that arose specifically in the common ancestor of all placental mammals - novel unpublished data generated by a PhD student co-supervised by Dr Forde and Dr O'Connell, as well as placental mammal phylogeny which has recently been published by Dr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.