Poster Presentation Society for Molecular Biology and Evolution Conference 2016

Unravelling marsupial X inactivation with shRNA knockdowns (#462)

Kim L McIntyre 1 , Shafagh A Waters 2 , Kevin V Morris 1 , Paul D Waters 1
  1. School of Biotechnology and Biomolecular Sciences, University Of New South Wales, Sydney, NSW, Australia
  2. The University Of New South Wales, Sydney, NSW, Australia

X chromosome inactivation is an epigenetic phenomenon occurring in eutherian and marsupial mammals to rectify imbalances in X-linked gene dosage. One X chromosome is transcriptionally silenced early in female embryonic development, which is then stably maintained in somatic tissue. In eutherians, the lncRNA Xist is transcribed from the future inactive X, coating it in cis to form the distinctive Xist cloud. Xist is known to interact with chromatin-modifying complexes. In marsupials, the independently evolved lncRNA Rsx silences the inactive X and forms the same distinctive cloud as Xist. Here I use RNA fluorescence in situ hybridization (RNA-FISH) to investigate how knocking down epigenetic modifiers changes Rsx localisation and reactivation of X-linked genes.