Unbiased identification of signal-activated transcription factors by barcoded synthetic tandem repeat promoter screening (BC-STAR-PROM)
| العنوان: | Unbiased identification of signal-activated transcription factors by barcoded synthetic tandem repeat promoter screening (BC-STAR-PROM) |
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| المؤلفون: | Pauline Gosselin, Ueli Schibler, Fabienne Fleury-Olela, Gianpaolo Rando |
| المصدر: | Genes & Development Genes Dev |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, Serum Response Factor, Base pair, genetic processes, Antineoplastic Agents, macromolecular substances, Biology, Vinblastine, Cell Line, 03 medical and health sciences, chemistry.chemical_compound, Mice, Tandem repeat, Depsipeptides, Serum response factor, Genetics, Genes, Synthetic, Animals, Humans, Luciferase, natural sciences, Promoter Regions, Genetic, Transcription factor, Cytoskeleton, Genetic Association Studies, Expression vector, fungi, Promoter, Molecular biology, female genital diseases and pregnancy complications, 3. Good health, Cell biology, 030104 developmental biology, chemistry, Genetic Techniques, Tandem Repeat Sequences, Gene Knockdown Techniques, DNA, Resource/Methodology, Developmental Biology, Signal Transduction, Transcription Factors |
| الوصف: | The discovery of transcription factors (TFs) controlling pathways in health and disease is of paramount interest. We designed a widely applicable method, dubbed barcorded synthetic tandem repeat promoter screening (BC-STAR-PROM), to identify signal-activated TFs without any a priori knowledge about their properties. The BC-STAR-PROM library consists of ∼3000 luciferase expression vectors, each harboring a promoter (composed of six tandem repeats of synthetic random DNA) and an associated barcode of 20 base pairs (bp) within the 3′ untranslated mRNA region. Together, the promoter sequences encompass >400,000 bp of random DNA, a sequence complexity sufficient to capture most TFs. Cells transfected with the library are exposed to a signal, and the mRNAs that it encodes are counted by next-generation sequencing of the barcodes. This allows the simultaneous activity tracking of each of the ∼3000 synthetic promoters in a single experiment. Here we establish proof of concept for BC-STAR-PROM by applying it to the identification of TFs induced by drugs affecting actin and tubulin cytoskeleton dynamics. BC-STAR-PROM revealed that serum response factor (SRF) is the only immediate early TF induced by both actin polymerization and microtubule depolymerization. Such changes in cytoskeleton dynamics are known to occur during the cell division cycle, and real-time bioluminescence microscopy indeed revealed cell-autonomous SRF–myocardin-related TF (MRTF) activity bouts in proliferating cells. |
| تدمد: | 1549-5477 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::547a7ed4daea2a55fff4015933796b79Test https://pubmed.ncbi.nlm.nih.gov/27601530Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....547a7ed4daea2a55fff4015933796b79 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15495477 |
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