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....
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| Format: | Final Year Project (FYP) |
| Language: | English |
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Nanyang Technological University
2020
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| Online Access: | https://hdl.handle.net/10356/139952 |
| _version_ | 1811687283280052224 |
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| author | Hew, Rachel Jia Yi |
| author2 | - |
| author_facet | - Hew, Rachel Jia Yi |
| author_sort | Hew, Rachel Jia Yi |
| collection | NTU |
| 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. |
| first_indexed | 2024-10-01T05:13:51Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/139952 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:13:51Z |
| publishDate | 2020 |
| publisher | Nanyang Technological University |
| record_format | dspace |
| spelling | ntu-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 |
| spellingShingle | Science::Biological sciences Hew, Rachel Jia Yi Optimisation of modRNA gene delivery and the FUCCI mouse model for cardiomyocyte proliferation |
| title | 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_short | 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 |
| topic | Science::Biological sciences |
| url | https://hdl.handle.net/10356/139952 |
| work_keys_str_mv | AT hewracheljiayi optimisationofmodrnagenedeliveryandthefuccimousemodelforcardiomyocyteproliferation |