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  • 學位論文

以聚合物作為載體運送小型干擾核醣核酸之研究

Study of the polymers as vectors for delivery of siRNA into cells.

指導教授 : 陳燕惠

摘要


核醣核酸干擾(RNA interference , RNAi )是近來發展的新技術,可以用來抑制特定的基因表現。許多研究利用RNAi來抑制病毒的複製,都得到不錯的效果,使得RNAi有相當高的潛力成為新的抗病毒藥物。 目前已經找到不少運送RNAi的載體,但是大多數效果很好的載體都有毒性過高的缺點,所以本研究針對低毒性的分子來尋找可能的載體。 Pluronic 是屬於非離子性的介面活性劑,毒性不高。我們利用Pluronic類的聚合物 F68 、F108與L44來運送GAPDH siRNA,實驗結果並沒有看到GAPDH受到抑制。接著我們利用合成以共價鍵連接Pluronic F68與分子量2000Da的PEI。實驗結果顯示以Pluronic F68-PEI (2K)來運送GAPDH siRNA,有觀察到GAPDH表現受到抑制,而對照組PEI (2K)則沒觀察到GAPDH受到抑制。另外使用Pluronic F108-PEI (2K)來運送siRNA的實驗中則沒有看到GAPDH表現受到抑制。

並列摘要


Small interference RNA ( siRNA ) is a new technology and is able to inhibit specific gene expression. SiRNA has been used to inhibit the replication of viruses in many studies and becomes a new potential antiviral agent. Many siRNA carriers have been developed. However, the more effective the carriers are, the higher toxicity they exert. Pluronic, a non-ionic surfactant with low toxicity, is considered as a carrier candidate in this study. There was no inhibition of GAPDH expression when Pluronic F68, F108 or L44 alone was used to deliver GAPDH siRNA. The Pluronic-grafted polyethyleneimine was then synthesized. The Pluronic F68-PEI (2K) moderately delivered GAPDH siRNA into BEAS-2B cells without detectable toxicity during transfection, while free PEI (2K) didn’t show the ability to deliver siRNA.

並列關鍵字

delivery RNAi siRNA micelle

參考文獻


1. Fire, A. et al., Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 1998. 391(6669): p.806-11
3. Elbashir, S.M., Lendeckel, W. and Tuschl, T., RNA interference is mediated by 21- and 22- nucleotide RNAs. Genes Dev, 2001. 15(2): p.188-200.
4. Chatterjee-Kishore, M. and Miller, CP., Exploring the sounds of silence: RNAi-mediated gene silencing for target identification and validation. Drug Discov Today, 2005. 10(22): p.1559-65.
5. Hannon, GJ., RNA interference. Nature, 2002. 418(6894): p.244-51.
6. Timmons, L. and Fire, A., Specific interference by ingested dsRNA. Nature, 1998. 395(6705): p.854.

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