What is Pol II ChIP seq?
PolII ChIP-seq can be used to profile genome-wide transcription activities by measuring PolII occupancies (e.g., ChIP-seq signals) over gene bodies. This powerful genome-wide transcription profiling method can capture active transcription events and global dynamic transcription changes in time-course experiments.
What is the function of RNA polymerase II?
RNA polymerase II (RNAP II and Pol II) is a multiprotein complex that transcribes DNA into precursors of messenger RNA (mRNA) and most small nuclear RNA (snRNA) and microRNA. It is one of the three RNAP enzymes found in the nucleus of eukaryotic cells.
Does RNA polymerase II unwind DNA?
RNA polymerase II allows unwinding and rewinding of the DNA and thus maintains a constant length of the transcription bubble. J Biol Chem.
How does Gro seq work?
GRO-Seq maps the binding sites of transcriptionally active RNA polymerase II (RNAPII). In this method, active RNAPII is allowed to run on in the presence of 5-bromouridine 5′-triphosphate (Br-UTP). RNAs are hydrolyzed and purified using beads coated with antibodies to 5-bromo-2-deoxyuridine (BrdU).
What are the functions of RNA polymerase I and II in eukaryotes?
RNA polymerase I (RNAPI) transcribes rRNA genes, RNA polymerase II (RNAPII) transcribes mRNA, miRNA, snRNA, and snoRNA genes, and RNA polymerase III (RNAPIII) transcribes tRNA and 5S rRNA genes. This is in contrast with prokaryotes where a single RNA polymerase is responsible for the transcription of all genes.
What is the function of RNA polymerase?
RNA polymerase (green) synthesizes RNA by following a strand of DNA. RNA polymerase is an enzyme that is responsible for copying a DNA sequence into an RNA sequence, duyring the process of transcription.
Can DNA polymerase unwind DNA?
The DNA polymerase requires the helicase to provide it with the ssDNA template for DNA synthesis. Similarly, the DNA polymerase serves to increase the unwinding rate of the helicase. In addition, the experiments show that the rate of DNA synthesis dictates the stimulated unwinding rate.
Where is RNA polymerase 2 found?
RNA polymerase II is located in the nucleus and synthesizes all protein-coding nuclear pre-mRNAs. Eukaryotic pre-mRNAs undergo extensive processing after transcription but before translation (Figure 1).
How is total mRNA measured in ChIP seq?
Cells were fixed, chromatin prepared and TranscriptionPath-Seq analysis performed using qPCR primers approximately 1500 bp 3′ of the transcription start site (TSS). Total mRNA was measured by preparing cDNA and designing primers that span splice junctions. Igf1r (A), Irs2 (B) and Egr1 (C) are all genes for which induced transcription was detected.
What kind of antibody is used for ChIP-seq?
Performing ChIP-Seq using an antibody against RNA pol II produces a genome-wide profile of gene transcription rates. ChIP-Seq was performed using chromatin from control and estrogen-treated MCF-7 cells and antibodies against RNA Pol II and the estrogen-inducible transcription factor SRC3.
How is ChIP seq used to detect down regulated genes?
TranscriptionPath also enables earlier detection and more dramatic differences for the down-regulated gene Slc2a2 (D) with the most dramatic decrease in transcription observed at 30 minutes compared to 3 hours for RT-qPCR measurements. ChIP-Seq is a widely-used method to detect transcription factor binding across the entire genome.