Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation

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....

Full description

Saved in:
Bibliographic Details
Main Author: Hew, Rachel Jia Yi
Other Authors: -
Format: Final Year Project
Language:English
Published: Nanyang Technological University 2020
Subjects:
Online Access:https://hdl.handle.net/10356/139952
Tags: Add Tag
No Tags, Be the first to tag this record!
Institution: Nanyang Technological University
Language: English
id sg-ntu-dr.10356-139952
record_format dspace
spelling sg-ntu-dr.10356-1399522023-02-28T18:08:25Z Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation Hew, Rachel Jia Yi - School of Biological Sciences Harvard University Department of Stem Cell and Regenerative Biology Richard T. Lee richard_lee@harvard.edu Science::Biological sciences 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. Bachelor of Science in Biological Sciences 2020-05-23T05:28:19Z 2020-05-23T05:28:19Z 2020 Final Year Project (FYP) https://hdl.handle.net/10356/139952 en application/pdf Nanyang Technological University
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
spellingShingle Science::Biological sciences
Hew, Rachel Jia Yi
Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
description 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.
author2 -
author_facet -
Hew, Rachel Jia Yi
format Final Year Project
author Hew, Rachel Jia Yi
author_sort Hew, Rachel Jia Yi
title Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
title_short Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
title_full Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
title_fullStr Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
title_full_unstemmed Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation
title_sort optimisation of modrna gene delivery and the fucci mouse model for cardiomyocyte proliferation
publisher Nanyang Technological University
publishDate 2020
url https://hdl.handle.net/10356/139952
_version_ 1759855409871454208