基因編碼FRET熒光傳感器，被設計用于在體外和活細胞中檢測不對稱二甲基化水平

2023年2月，中國科學技術大學基礎醫學院生命科學與醫學部中國科學院先天免疫與慢性病重點實驗室合肥微尺度物理科學國家實驗室;中國科學技術大學**附屬醫院(Hefei National Laboratory for Physical Sciences at Microscale, the CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China；The First Afliated Hospital of University of Science and Technology of China, Hefei 230001, China) Xuan Sun老師研究團隊在《Analytical and Bioanalytical Chemistry》上發表論文：

“A gene-encoded FRET fluorescent sensor designed for detecting asymmetric dimethylation levels in vitro and in living cells”

“基因編碼FRET熒光傳感器，被設計用于在體外和活細胞中檢測不對稱二甲基化水平”

Abstract：

Arginine methylation is involved in many important biological processes. PRMT1 is a major arginine methyltransferase in mammalian cells and is highly conserved in eukaryotes. It catalyzes the methylation of various of substrates, including histones, and PRMT1 has been reported to be overexpressed in many cancers, indicating that it is a potential therapeutic target. No tool for efficient methylation level detection in living cells has been available to date. In this work, we designed and constructed a gene-encoded fluorescence resonance energy transfer (FRET) fluorescent sensor for detecting dimethylation levels in living cells and evaluated its functional efficiency both in vitro and in living cells. Both site-directed mutagenesis and PRMT1 inhibition experiments verified that the fluorescent sensor responded to changes in PRMT1 activity and to different PRMT1-induced methylation levels in vitro. Finally, we verified that this optimized methyl sensor responded sensitively to changes in methylation levels in living cells by overexpressing and inhibiting PRMT1, which makes it a useful tool for real-time imaging of arginine methylation. As a new tool for detecting arginine dimethylation levels in living cells, the designed FRET sensor is very important for posttranslational studies and may show a wide range of applications.

摘要：

精氨酸甲基化參與了許多重要的生物過程。PRMT1是哺乳動物細胞中主要的精氨酸甲基轉移酶，在真核生物中高度保守。它催化包括組蛋白在內的多種底物的甲基化，據報道，PRMT1在許多癌癥中過表達，這表明它是一個潛在的治療靶點。迄今為止，還沒有有效的活細胞甲基化水平檢測工具。

在這項工作中，科研人員設計并構建了一個基因編碼熒光共振能量轉移(FRET)熒光傳感器，用于檢測活細胞中的二甲基化水平，并評估了其在體外和活細胞中的功能效率。位點定向誘變和PRMT1抑制實驗都證實了熒光傳感器對PRMT1活性的變化和不同PRMT1誘導的體外甲基化水平有響應。最后，科研人員驗證了這種優化的甲基傳感器通過過表達和抑制PRMT1對活細胞中甲基化水平的變化做出敏感的反應，這使其成為精氨酸甲基化實時成像的有用工具。作為一種檢測活細胞中精氨酸二甲基化水平的新工具，所設計的FRET傳感器在翻譯后研究中具有重要意義，具有廣泛的應用前景。

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