Establishing two transgenic mouse cell lines for future studies on X chromosome inactivation : a red Xi in live cells and a silenced GFP on the Xi.

• Main Author: Ong, Norbert Ha How.
• Other Authors: School of Biological Sciences
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/53818
• Institution: Nanyang Technological University

X chromosome inactivation (XCI) is a classic example of dosage compensation in the female eutherians. XCI is defined as a process in which one of the X chromosomes in a XX female cell is stochastically chosen to be inactivated. Consequently, a balance in X-linked gene dosages between female and male cells is achieved. Despite XCI has been actively studied for more than 50 years, the exact process of XCI remains elusive. Hence, the aim of this project was to generate two female mouse transgenic cell lines that could be used for studying XCI. In our study, the first cell line was generated using lipo-transfection method. Here, female mouse fibroblast cells were stably transfected with plasmids that were engineered with macroH2A–mCherry transgene. Our results showed that, if expressed, this fusion protein would accumulate on the inactive X chromosome (Xi) and thus showing a red Xi. Additionally, the second cell line was generated using fluorescence-activated cell sorting (FACS) method. FACS was used to isolate Xi–GFP and Xa–GFP cells from an existing transgenic female mouse strain that was engineered with an X-linked GFP gene. In conclusion, macroH2A–mCherry transgenic cell line and X–GFP cell lines were valuable tools that could be used to further investigate the process of XCI.