Site directed mutagenesis of miRNA binding site on the 3′-UTR of choline kinase alpha gene
MicroRNA largely controls gene expression by attaching to messenger RNA (mRNA) in the cell cytoplasm. Instead of being promptly translated into a protein, the targeted mRNA will either be destroyed and its components recycled, or it will be retained and translated later. Choline kinase alpha (chk...
| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2022
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| Subjects: | |
| Online Access: | http://eprints.usm.my/58370/ http://eprints.usm.my/58370/1/Bnisallama-24%20pages.pdf |
| _version_ | 1848883880641167360 |
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| author | Bnisallama, Faisal Fayez Saffah |
| author_facet | Bnisallama, Faisal Fayez Saffah |
| author_sort | Bnisallama, Faisal Fayez Saffah |
| building | USM Institutional Repository |
| collection | Online Access |
| description | MicroRNA largely controls gene expression by attaching to messenger RNA (mRNA) in
the cell cytoplasm. Instead of being promptly translated into a protein, the targeted mRNA
will either be destroyed and its components recycled, or it will be retained and translated
later. Choline kinase alpha (chka) overexpression is a clinical sign of diseased tissues and
malignant cells. MicroRNAs (miRNAs) are effective posttranscriptional regulators of
gene. Studies by our team showed that these three miRNAs (miR-876-5p, miR-367-3p
and miR-32-5p) downregulated the expression of chka gene. However, the binding sites
of these miRNAs on the 3'-UTR of the chka gene have not been verified. This study aimed
to mutate the binding sites of these miRNAs for subsequent verification by a luciferase
assay. In this study, we performed PCR site-directed mutagenesis on the miR-367-3p
(GAAGCAGAAAT ATAGTGCAATA) from nucleotides (nt) 1817-1825 binding sites in
chka and miR-876-5p (GAG TGTAGCTGTG AAATCCA) binding site from nucleotides
(nt) 2573-2581 binding sites and verified the mutation by DNA sequencing. In vitro work,
after mutagenesis step, the PCR products were sent for sequencing, however the results
were not satisfactory. |
| first_indexed | 2025-11-15T18:57:50Z |
| format | Thesis |
| id | usm-58370 |
| institution | Universiti Sains Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T18:57:50Z |
| publishDate | 2022 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | usm-583702023-05-15T02:39:56Z http://eprints.usm.my/58370/ Site directed mutagenesis of miRNA binding site on the 3′-UTR of choline kinase alpha gene Bnisallama, Faisal Fayez Saffah QK900-989 Plant ecology MicroRNA largely controls gene expression by attaching to messenger RNA (mRNA) in the cell cytoplasm. Instead of being promptly translated into a protein, the targeted mRNA will either be destroyed and its components recycled, or it will be retained and translated later. Choline kinase alpha (chka) overexpression is a clinical sign of diseased tissues and malignant cells. MicroRNAs (miRNAs) are effective posttranscriptional regulators of gene. Studies by our team showed that these three miRNAs (miR-876-5p, miR-367-3p and miR-32-5p) downregulated the expression of chka gene. However, the binding sites of these miRNAs on the 3'-UTR of the chka gene have not been verified. This study aimed to mutate the binding sites of these miRNAs for subsequent verification by a luciferase assay. In this study, we performed PCR site-directed mutagenesis on the miR-367-3p (GAAGCAGAAAT ATAGTGCAATA) from nucleotides (nt) 1817-1825 binding sites in chka and miR-876-5p (GAG TGTAGCTGTG AAATCCA) binding site from nucleotides (nt) 2573-2581 binding sites and verified the mutation by DNA sequencing. In vitro work, after mutagenesis step, the PCR products were sent for sequencing, however the results were not satisfactory. 2022-09 Thesis NonPeerReviewed application/pdf en http://eprints.usm.my/58370/1/Bnisallama-24%20pages.pdf Bnisallama, Faisal Fayez Saffah (2022) Site directed mutagenesis of miRNA binding site on the 3′-UTR of choline kinase alpha gene. Masters thesis, Universiti Sains Malaysia. |
| spellingShingle | QK900-989 Plant ecology Bnisallama, Faisal Fayez Saffah Site directed mutagenesis of miRNA binding site on the 3′-UTR of choline kinase alpha gene |
| title | Site directed mutagenesis of miRNA binding site
on the 3′-UTR of choline kinase alpha gene |
| title_full | Site directed mutagenesis of miRNA binding site
on the 3′-UTR of choline kinase alpha gene |
| title_fullStr | Site directed mutagenesis of miRNA binding site
on the 3′-UTR of choline kinase alpha gene |
| title_full_unstemmed | Site directed mutagenesis of miRNA binding site
on the 3′-UTR of choline kinase alpha gene |
| title_short | Site directed mutagenesis of miRNA binding site
on the 3′-UTR of choline kinase alpha gene |
| title_sort | site directed mutagenesis of mirna binding site
on the 3′-utr of choline kinase alpha gene |
| topic | QK900-989 Plant ecology |
| url | http://eprints.usm.my/58370/ http://eprints.usm.my/58370/1/Bnisallama-24%20pages.pdf |