1. Transient transfection of mammalian cells using a violet diode laser
- Author
-
Maria Leilani Torres-Mapa, Liselotte Angus, Martin Ploschner, Kishan Dholakia, and Frank J. Gunn-Moore
- Subjects
MST1 ,Biomedical Engineering ,CHO Cells ,Biology ,Transfection ,Biomaterials ,Ezrin ,Cricetulus ,Radixin ,Cricetinae ,Animals ,Humans ,RNA, Small Interfering ,Hippo signaling pathway ,FERM domain ,Kinase ,Lasers ,Cell Membrane ,Equipment Design ,Atomic and Molecular Physics, and Optics ,Hedgehog signaling pathway ,Electronic, Optical and Magnetic Materials ,Cell biology ,Equipment Failure Analysis ,HEK293 Cells ,Semiconductors ,Cancer research ,Phosphorylation - Abstract
The Salvador/Warts/Hippo (Hippo) pathway defines a novel signalling cascade regulating cell contact inhibition, organ size control, cell growth, proliferation, apoptosis and cancer development in mammals. The Hippo pathway was initially utilised in D. melanogaster, where the Expanded protein acts in the Hippo signalling cascade to control organ size. Willin is the proposed human orthologue of Expanded and the aim of this thesis is to investigate whether willin can activate the mammalian Hippo signalling pathway. Ectopic willin expression causes an increase in phosphorylation of the core Hippo signalling pathway components MST1/2, LATS1 and YAP, an effect which can be antagonised by ezrin. In MCF10A cells, willin over-expression antagonises a YAP-induced epithelial-to-mesenchymal transition via the N- terminal FERM (Four-point-one Ezrin Radixin Moesin) domain of willin. Preliminary results show that willin is expressed within the sciatic nerve of rat and mice, and within the neuromast cells in the zebrafish; suggesting that willin and the Hippo pathway may play a vital role in the developmental regulation within the peripheral nervous system. To conclude, willin influences Hippo signalling activity by activating the core Hippo pathway kinase cassette in mammalian cells.
- Published
- 2010