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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 詹世鵬(Shih-Peng Chan) | |
| dc.contributor.author | Wei-Lin Lin | en |
| dc.contributor.author | 林薇琳 | zh_TW |
| dc.date.accessioned | 2022-11-25T06:34:55Z | - |
| dc.date.copyright | 2021-11-09 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-09-29 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82281 | - |
| dc.description.abstract | 秀麗隱桿線蟲（Caenorhabditis elegans）的生長與發育主要由一連串具有時間性的基因所控制，這群基因被稱為異時性基因（Heterochronic genes），其中包括特定的微小核糖核酸（micro RNAs）以及RNA結合蛋白。其中，具有高度保留性的微小核糖核酸let-7便是影響到線蟲由幼蟲進入到成蟲階段的一個重要因子，let-7的缺失會使線蟲在成蟲時出現接縫細胞（epithelial stem-like seam cell）重複分裂、生殖孔爆裂等性狀。在人類中，let-7的缺失同樣會影響細胞的分化，導致癌症的發生。let-7在線蟲中主要由LIN-28所調控，並會抑制下游lin-41的表現。而lin-41的降低會使LIN-29開始表現，因此let-7的缺失會間接導致LIN-29的表現被抑制。 Musashi蛋白是一種RNA結合蛋白，主要被發現於幹細胞中，含量過高時與癌症的發生有很高的相關性。先前有研究指出，在小鼠胚胎幹細胞中，MSI-1可能會透過結合LIN-28蛋白間接調控下游let-7 miRNA的生成，讓我們對於MSI-1在線蟲中與let-7的關聯性與異時性途徑產生興趣。為了探討MSI-1對於異時性途徑的影響，我們利用會同時表現兩種由不同3′ UTR調控的雙色螢光報告子的線蟲，GFP由lin-41 3′ UTR所控制，而mCherry由不受let-7影響的unc54 3′ UTR所調控，藉由螢光的變化去觀察利用小核糖核酸干擾技術去降低MSI-1表現時是否對let-7功能造成影響。可惜的是，就我們初步的觀察，MSI-1的下降並不會對於蟲的let-7對lin-41 3′ UTR reporter 的調控表現量以及其變異的性狀造成影響。 與此同時，我們也利用雙色螢光報告子線蟲在具有潛力的基因中，利用篩選新的研究目標。透過雙色螢光蟲的GFP變化，我們發現有兩個基因ani-1和mix-1表現量可能會影響到let-7對於lin-41 3′ UTR的控制，但在利用北方墨點法做檢查以後，發現它們與let-7表現量沒有關係。但對於它們是否透過其他的途徑去影響到seam cells的變化，仍有待未來進一步探討。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T06:34:55Z (GMT). No. of bitstreams: 1 U0001-2709202119542800.pdf: 4620206 bytes, checksum: b38f03b74fbdd1c1ede7c44d7fdd7b85 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "口試委員審定書 I 誌謝 II 中文摘要 III Abstract IV Contents VI Introduction 1 The overview of miRNAs 1 The let-7 miRNA in C. elegans 2 lin-41, a downstream gene of let-7 3 MSI-1, previously thought to be involved in let-7-related regulation 4 Other candidates tested in this work that may affect let-7 function 5 Materials and methods 8 Strains 8 Culture 9 RNA interference 9 Bursting Assay 10 Genomic DNA extraction 10 RNA isolation 11 Protein extraction 11 Construction of RNAi plasmids 12 Vector Preparation 12 Insert Preparation 12 Ligation 12 Western Blot 13 Northern Blot 14 Results 16 Knockdown of msi-1 would not inhibit the mutant phenotype of low expression of let-7. 16 The dual fluorescence reporter inserted into BRC0566 is single copy and complete 17 The transgene worms SPN401 and SPN405 could be used as a detection tool for observing the lin-41 3′ UTR control of let-7 by monitoring the fluorescent change 18 Knockdown of msi-1 does not affect let-7 repressing lin-41 3′ UTR 20 Screening potential candidates by observing SPN497 21 Knockdown of ani-1 and mix-1 would not affect the expression of let-7 in C. elegans. 22 Discussion 23 The application of dual-fluorescent worms 23 The potential targets from let-7 interactome 24 Figures 27 Figure 1 27 Figure 2 29 Figure 3 31 Figure 4 35 Figure 5 38 Figure 6 40 Figure 7 43 Figure 8 46 Tables 47 Table 1 47 Table 2 50 References 51 Appendix 55 Appendix 1 55 Appendix 2 56 Appendix 3 57 Appendix 4 58 Appendix 5 60 Appendix 6 61 Appendix 7 62" | |
| dc.language.iso | en | |
| dc.subject | mix-1 | zh_TW |
| dc.subject | 微小核醣核酸 | zh_TW |
| dc.subject | let-7 | zh_TW |
| dc.subject | lin-41 | zh_TW |
| dc.subject | MSI-1 | zh_TW |
| dc.subject | ani-1 | zh_TW |
| dc.subject | micro RNAs | en |
| dc.subject | ani-1 | en |
| dc.subject | MSI-1 | en |
| dc.subject | lin-41 | en |
| dc.subject | let-7 | en |
| dc.subject | mix-1 | en |
| dc.title | 利用雙色螢光報導子線蟲探討let-7/lin-41路徑中的可能的調控因子 | zh_TW |
| dc.title | Using a dual-fluorescence 3′ UTR reporter to investigate putative regulators in the C. elegans let-7/lin-41 pathway | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 顏柏勳(Hsin-Tsai Liu),陳俊豪(Chih-Yang Tseng) | |
| dc.subject.keyword | 微小核醣核酸,let-7,lin-41,MSI-1,ani-1,mix-1, | zh_TW |
| dc.subject.keyword | micro RNAs,let-7,lin-41,MSI-1,ani-1,mix-1, | en |
| dc.relation.page | 62 | |
| dc.identifier.doi | 10.6342/NTU202103413 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2021-09-29 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| dc.date.embargo-lift | 2026-08-22 | - |
| 顯示於系所單位： | 微生物學科所 | |
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