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Title: Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
Authors: Hew, Rachel Jia Yi
Keywords: Science::Biological sciences
Issue Date: 2020
Publisher: Nanyang Technological University
Abstract: The human heart is known to have limited regenerative capacity, with minimal proliferation of pre-existing cardiomyocytes. After a myocardial infarction, extensive cardiomyocyte death occurs, resulting in heart failure. Thus, efforts have been made to identify remaining proliferative cardiomyocytes. Fluorescence Ubiquitination-based Cell Cycle Indicator (FUCCI) is one method that is able to discern the cell cycle phase of an arrested cell. When the cell is arrested at the G1/S transition, the nucleus appears yellow, while in G2 and mitosis phases, the nucleus appears red. In the S phase, the nucleus appears green, indicating that the cell is replicating DNA for proliferation. In this project, we aim to use FUCCI to monitor cell cycle progression of cardiomyocytes transfected with different modified RNA (modRNA) molecules to identify potential proliferative gene targets. After optimizing modRNA production to ensure efficient transfection, we used these modRNA molecules to transfect isolated FUCCI cardiomyocytes. From this study, we concluded that high quality modRNA molecules were successfully made. However, proliferative capabilities after transfection of the cardiomyocytes have yet to be determined. Further studies will be required to improve the transfection protocol and data analysis methods to investigate the FUCCI system’s reliability as a proliferative marker.
Fulltext Permission: restricted
Fulltext Availability: With Fulltext
Appears in Collections:SBS Student Reports (FYP/IA/PA/PI)

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