Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA

Pseudouridylation is the most abundant type of Ribonucleic acid (RNA) modification where uracil is converted into pseudouridine (Ψ). The 5’ end of U1 small nuclear RNA (snRNA) contains two evolutionary conserved pseudouridines, Ψ5 and Ψ6. It is still a challenge to understand the splicing roles of Ψ...

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Main Author: Tan, Darren Wei
Other Authors: Francesc Xavier Roca Castella
Format: Final Year Project
Language:English
Published: 2019
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Online Access:http://hdl.handle.net/10356/77190
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-771902023-02-28T18:01:46Z Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA Tan, Darren Wei Francesc Xavier Roca Castella School of Biological Sciences DRNTU::Science::Biological sciences Pseudouridylation is the most abundant type of Ribonucleic acid (RNA) modification where uracil is converted into pseudouridine (Ψ). The 5’ end of U1 small nuclear RNA (snRNA) contains two evolutionary conserved pseudouridines, Ψ5 and Ψ6. It is still a challenge to understand the splicing roles of Ψ5 and Ψ6 in helping U1 to recognise thousands of 5’ splice site (5’ss) variants. Hence, our project is to characterize the effects of removing the Ψ5 and Ψ6 in U1 snRNA on splicing changes via scaRNAs ACA47 and U109 Knockout (KO) which act as guide RNAs for pseudouridylation to occur. Differentially expressed genes (DEGs) and time-course of rescue plasmids experiments were conducted to investigate if Ψ5 and Ψ6 affect the gene expression after ACA47 and U109 KO. Minigene experiments were also performed to observe the splicing effects of Ψ5 and Ψ6. Results showed that indeed Ψ5 and Ψ6 do affect the gene expression and longer transfection time would increase the percentage of rescue DEGs. The splicing pattern of Ψ5 and Ψ6 can also cause an indirect effect of gene regulation. In summary, our findings are useful for better understanding of the 5’ splice site (5’ss)/U1 recognition. Bachelor of Science in Biological Sciences 2019-05-15T07:04:35Z 2019-05-15T07:04:35Z 2019 Final Year Project (FYP) http://hdl.handle.net/10356/77190 en Nanyang Technological University 30 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences
spellingShingle DRNTU::Science::Biological sciences
Tan, Darren Wei
Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA
description Pseudouridylation is the most abundant type of Ribonucleic acid (RNA) modification where uracil is converted into pseudouridine (Ψ). The 5’ end of U1 small nuclear RNA (snRNA) contains two evolutionary conserved pseudouridines, Ψ5 and Ψ6. It is still a challenge to understand the splicing roles of Ψ5 and Ψ6 in helping U1 to recognise thousands of 5’ splice site (5’ss) variants. Hence, our project is to characterize the effects of removing the Ψ5 and Ψ6 in U1 snRNA on splicing changes via scaRNAs ACA47 and U109 Knockout (KO) which act as guide RNAs for pseudouridylation to occur. Differentially expressed genes (DEGs) and time-course of rescue plasmids experiments were conducted to investigate if Ψ5 and Ψ6 affect the gene expression after ACA47 and U109 KO. Minigene experiments were also performed to observe the splicing effects of Ψ5 and Ψ6. Results showed that indeed Ψ5 and Ψ6 do affect the gene expression and longer transfection time would increase the percentage of rescue DEGs. The splicing pattern of Ψ5 and Ψ6 can also cause an indirect effect of gene regulation. In summary, our findings are useful for better understanding of the 5’ splice site (5’ss)/U1 recognition.
author2 Francesc Xavier Roca Castella
author_facet Francesc Xavier Roca Castella
Tan, Darren Wei
format Final Year Project
author Tan, Darren Wei
author_sort Tan, Darren Wei
title Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA
title_short Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA
title_full Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA
title_fullStr Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA
title_full_unstemmed Characterization of the splicing functions of the two pseudouridines Ψ5 and Ψ6 in U1 small nuclear RNA
title_sort characterization of the splicing functions of the two pseudouridines ψ5 and ψ6 in u1 small nuclear rna
publishDate 2019
url http://hdl.handle.net/10356/77190
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