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Stitzinger, S.

Publications and source records attributed to Stitzinger, S..

2 recordsLinked to original sources

Non-coding RNA controls long-range enhancer-promoter interactions

Transcription regulation depends on interactions between enhancers and gene promoters, which often occur over large genomic distances. The molecular drivers of long-range enhancer-promoter interactions remain poorly understood. Enhancers themselves are known to be transcribed into non-coding RNAs, but the function of these transcripts is unclear. Here, we use acute genetic and chemical perturbations of transcription elongation and termination to study the role of non-coding RNA in enhancer-promoter interactions. By integrating high-resolution 3D genome mapping, transcriptomics, proteomics, biochemistry and live-cell super-resolution imaging in these perturbation contexts, we demonstrate that a strong increase in RNA molecules at enhancers disrupts their long-range interactions with gene promoters, whereas decreased RNA levels lead to stabilization of these interactions. Using separation-of-function perturbations, we show that this regulatory function of RNA is independent from the process of transcription and the proteins recruited to initiate transcription. Functionally, we show that RNA-mediated disruption of enhancer-promoter interactions is associated with reduced transcription of target genes, indicating a regulatory feedback mechanism. Together, our results uncover an additional regulatory principle of enhancer-promoter interactions mediated by non-coding RNA.

genomics↗

Continuous muscle, glial, epithelial, neuronal, and hemocyte cell lines for Drosophila research

Expression of activated Ras, RasV12, provides Drosophila cultured cells with a proliferation and survival advantage that simplifies the generation of continuous cell lines. Here we used lineage restricted RasV12 expression to generate continuous cell lines of muscle, glial, and epithelial cell type. Additionally, cell lines with neuronal and hemocyte characteristics were isolated by cloning from cell cultures established with broad RasV12 expression. Differentiation with the hormone ecdysone caused maturation of cells from mesoderm lines into active muscle tissue and enhanced dendritic features in neuronal-like lines. Transcriptome analysis showed expression of key cell-type specific genes and the expected alignment with single cell sequencing data in several cases. Overall, the technique has produced in vitro cell models with characteristics of glia, epithelium, muscle, nerve, and hemocyte. The cells and associated data are available from the Drosophila Genomic Resource Center.

cell biology↗