Alternative Splicing Caused by Iron Deficiency (I)

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Project title

Code/計畫編號

NSC102-2633-B019-001

Translated Name/計畫中文名

鐵離子缺乏是否會造成選擇性剪接( I )

Project Coordinator/計畫主持人

Wen-Shyong Tzou

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Bioscience and Biotechnology

Website

https://www.grb.gov.tw/search/planDetail?id=3169039

Year

2013

Start date/計畫起

01-10-2013

Expected Completion/計畫迄

30-09-2014

Bugetid/研究經費

550千元

ResearchField/研究領域

生物技術(農)

"鐵參與多項生物反應與促成多項生物功能，例如氧氣的運輸、氧化還原反應、細胞運輸與能量製造、免疫系統、神經系統與酵素的輔基等。我們從數十萬 exon與 intron邊界序列做一詳細檢測，我們發現人類與斑馬魚基因體中，有很多 iron-response element (IRE)出現:在人類有 23個個基因、斑馬魚有 12個基因的 exon與 intron邊界中有 IRE存在，這些 potential IRE都跨坐在 intron與 exon邊界上。我們提出一個假說:在鐵離子缺乏的情況下，造成 IRP 穩定，與 pre-mRNA 的 intron-exon邊界處結合，造成的選擇性剪接，會對於蛋白質結構與功能產生影響。本研究以人類細胞與斑馬魚胚胎為實驗模型，首先以 paired-end RNAseq 技術探究在鐵離子缺乏的情況下是否會有不同的基因選擇性剪接，繼而以不同的實驗設計 (IRP與 pre-mRNA結合、IRP與 splicing factor結合、cassette exon的 splicing junction 對 IRE的依賴性)確定在 exon-intron boundary存在的 IRE直接參與 alternative splicing。本研究將可以證實鐵的恆定性與基因的剪切以及蛋白質功能轉換的直接連結，更進一步對於地球工程中以鐵作為大量的鐵刺激浮游生物生長，促進固碳作用的方法可能產生的副作用提出科學證據。" "Iron is required for the surviving of organisms by contributing to multiple biological functions, such as the transport of oxygen, oxidation-reduction reaction, cellular transport, energy production, immune system, nervous system and act as enzyme cofactor. We examined millions of exon-intron boundaries of human and zebrafish genomes and we discovered 23 genes of human and 12 genes of zebrafish contained the iron regulatory element (IRE) in the exon-intron boundary. We hypothesized that binding of IRE binding protein (IRP) on IRE could lead to the change of spliceosome and skipping of several exons during splicing could take place. The new protein product through alternative splicing could contribute the repertoires of protein functions. In this research we will use human cell line and zebrafish embryo as a model system to test our hypothesis. We will conduct next generation sequencing of RNA (paired-end RNA-seq) to investigate the changes of exon choice of expressed genes upon the iron deficiency. Furthermore, we employed three methods to demonstrate the involvement of IRE on exon choice (IRP-pre-mRNA binding, IRP-splicing factor binding, and the dependence of alternative splicing on IRE by cassette exon). This study will be able to, for the first time, provide the direct link among iron homeostasis, gene splcing and protein function. Moreover, this reseach will also deliver the scientific evidence of the possible side effect of European Iron Fertilization Experiment as an effort of carbon sequestration."

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Alternative Splicing Caused by Iron Deficiency (I)

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