在哺乳類細胞中藉由Qβ類病毒顆粒快捷運輸平臺過度表現粒線體衍生性多肽(MOTS-c)

Abstract

從粒線體12S核糖體核糖核酸之編碼中發現一段粒線體衍生性多肽：MOTS-c (一段位於粒線體12S核糖體核糖核酸之開放讀序框架)。在細胞層級中，MOTS-c同時被發現可調節胰島素敏感性及代謝恆定性。有賴生物科技與醫學的發展，對於以奈米尺度在生物體內運輸小分子之快捷運輸平臺研究日趨重要。我們實驗室業已發展一套以類病毒顆粒為轉染媒介的核糖核酸運輸系統。類病毒顆粒是一種可將蛋白質或核糖核酸包裹在內的生物性腔室。在本研究中，我使用一種從Qβ核糖核酸噬菌體衍生而來的重組型20面體奈米微粒：Qβ類病毒顆粒直接包裹MOTS-c之信使核糖核酸(QβMOTS-c 類病毒顆粒)。接著以QβMOTS-c 類病毒顆粒處理HeLa細胞使其產生MOTS-c多肽。我們發展數種生化分析方法以鑑定產物，像是西方墨點法以及競爭型酶聯免疫分析法。我們的目標是生產在生物體層級中能有效調節代謝恆定性的MOTS-c多肽。

MOTS-c (mitochondrial open reading frame of the 12S rRNA-c) is a mitochondrial-derived peptide that is found to be encoded within the mitochondrial 12S rRNA. The MOTS-c is also found to regulate insulin sensitivity and metabolic homeostasis at the cellular level. Convenient platforms for delivering small molecules in vivo in nanoscale of interest are becoming more desirable as the development of biotechnology and medicine. An RNA-delivery system utilizes VLPs (virus-like particles) as a transfection agent is developed prior in our lab. The VLP is a biological compartment that allows one to pack with proteins or RNAs. Here I use the Qβ VLP, a recombinant icosahedral nanoparticle derived from the RNA bacteriophage Qβ, to directed pack the MOTS-c mRNA (QβMOTS-c VLP). The QβMOTS-c VLP is then used to produce the MOTS-c peptide in vivo in the HeLa cells. Several biochemistry assays such as western blot and the competitive enzyme-linked immunosorbent assay (ELISA) are developed to identify the product. Our goal is to produce an effective MOTS-c peptide to functionally regulate metabolic homeostasis at the organismal level.