A new strategy for screening novel functional genes involved in reduction of lipid droplet accumulation

IF 5 3区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

BioFactors Pub Date : 2023-11-20 DOI:10.1002/biof.2019

Ryuto Maruyama, Yasuhiro Kudo, Tomoyasu Sugiyama

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引用次数: 0

Abstract

Lipid droplets (LDs) are organelles that store excess lipids and provide fatty acids for energy production during starvation. LDs are also essential for cellular maintenance, but excessive accumulation of LDs triggers various cancers in addition to metabolic diseases such as diabetes. In this study, we aimed to develop a strategy to identify new genes that reduces accumulation of LDs in cancer cells using an RNA interference (RNAi) screening system employing artificial sequence-enriched shRNA libraries. Monitoring LDs by fluorescent activated cell sorting, the subsequently collected cumulative LDs cells, and shRNA sequence analysis identified a clone that potentially functioned to accumulate LDs. The clone showed no identical sequence to human Refseq. It showed very similar sequence to seven genes by allowing three mismatches. Among these genes, we identified the mediator complex subunit 6 (MED6) gene as a target of this shRNA. Silencing of MED6 led to an increase in LD accumulation and expression of the marker genes, PLIN2 and DGAT1, in fatty cells. MED6 is a member of the mediator complex that regulates RNA polymerase II transcription through transcription factor II. Some mediator complexes play important roles in both normal and pathophysiological transcription processes. These results suggest that MED6 transcriptionally regulates the genes involved in lipid metabolism and suppresses LD accumulation.

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筛选参与减少脂滴积累的新功能基因的新策略。

脂滴(ld)是储存多余脂质并在饥饿时提供脂肪酸以产生能量的细胞器。ld对细胞的维持也是必不可少的，但ld的过度积累除了引发糖尿病等代谢疾病外，还会引发各种癌症。在这项研究中，我们的目标是开发一种策略，利用RNA干扰(RNAi)筛选系统，利用人工序列富集shRNA文库，识别减少癌细胞中LDs积累的新基因。通过荧光活化细胞分选监测ld，随后收集累积ld细胞，并通过shRNA序列分析确定了一个可能具有积累ld功能的克隆。克隆的基因序列与人类的Refseq基因序列不相同。通过允许三次错配，它显示出与七个基因非常相似的序列。在这些基因中，我们确定了介质复合体亚基6 (MED6)基因作为该shRNA的靶标。MED6的沉默导致脂肪细胞中LD积累和标记基因PLIN2和DGAT1的表达增加。MED6是通过转录因子II调节RNA聚合酶II转录的中介复合体的成员。一些介质复合物在正常和病理生理转录过程中都起着重要作用。这些结果表明MED6通过转录调控脂质代谢相关基因，抑制LD积累。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

BioFactors 生物-内分泌学与代谢

CiteScore

11.50

自引率

3.30%

发文量

审稿时长

6-12 weeks

期刊介绍： BioFactors, a journal of the International Union of Biochemistry and Molecular Biology, is devoted to the rapid publication of highly significant original research articles and reviews in experimental biology in health and disease. The word “biofactors” refers to the many compounds that regulate biological functions. Biological factors comprise many molecules produced or modified by living organisms, and present in many essential systems like the blood, the nervous or immunological systems. A non-exhaustive list of biological factors includes neurotransmitters, cytokines, chemokines, hormones, coagulation factors, transcription factors, signaling molecules, receptor ligands and many more. In the group of biofactors we can accommodate several classical molecules not synthetized in the body such as vitamins, micronutrients or essential trace elements. In keeping with this unified view of biochemistry, BioFactors publishes research dealing with the identification of new substances and the elucidation of their functions at the biophysical, biochemical, cellular and human level as well as studies revealing novel functions of already known biofactors. The journal encourages the submission of studies that use biochemistry, biophysics, cell and molecular biology and/or cell signaling approaches.