| Autor: |
Steffi, Klier, Sarah, Farmer, Robertus A M, de Bruin |
| Rok vydání: |
2014 |
| Předmět: |
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| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 1170 |
| ISSN: |
1940-6029 |
| Popis: |
G1-S transcriptional control involves the coordination of the expression of a large set of co-regulated genes as a function of cell cycle progression (Bertoli et al., Nat Rev Mol Cell Biol 14:518-528, 2013). Confining transcription to the G1 phase of the cell cycle requires the regulation of specific transcription factor activity through either co-factors or regulation of promoter DNA binding. Therefore, the analysis of G1-S transcriptional control involves cell cycle synchronization and monitoring cell cycle synchrony, in order to establish DNA binding of G1-S transcription factors to G1-S promoters and to investigate changes in gene expression during the different phases of the cell cycle. Here, we describe a cell cycle synchrony method and ways to monitor synchrony. We also describe a chromatin immunoprecipitation (ChIP) method to locate G1-S transcription factor components to promoters and a quantitative PCR (qPCR) protocol to determine gene expression. Defining the binding dynamics of G1-S transcription factors and changes in gene expression during the cell cycle should provide new insights into the mechanism that control G1-S transcription and will allow for investigation of the biological relevance of confining gene expression to G1. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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