A new strategy for screening novel functional genes involved in reduction of lipid droplet accumulation.
Ryuto MaruyamaYasuhiro KudoTomoyasu SugiyamaPublished in: BioFactors (Oxford, England) (2023)
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.
Keyphrases
- genome wide identification
- fatty acid
- transcription factor
- genome wide
- induced apoptosis
- bioinformatics analysis
- cell cycle arrest
- endothelial cells
- type diabetes
- single cell
- cardiovascular disease
- genome wide analysis
- dna methylation
- amino acid
- poor prognosis
- gene expression
- adipose tissue
- stem cells
- physical activity
- cell death
- metabolic syndrome
- endoplasmic reticulum stress
- weight gain
- insulin resistance
- single molecule
- long non coding rna
- binding protein
- induced pluripotent stem cells