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Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events
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JoVE Journal Genetics
Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events

DOI:
10.3791/59497-v

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10:59 min

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May 13, 2019

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, ,
1Stowers Institute for Medical Research, 2Department of Biochemistry and Molecular Biology,Kansas University Medical Center

Chapters

  • 00:04Title
  • 00:31Assembly of Artificial Elongation Complexes
  • 04:36Walking Pol II to make 23, 25, and 29 Nucleotide Transcripts
  • 06:30Using Artificial Elongation Complexes to Assay Co-transcriptional Capping
  • 09:01Results: Artificial Elongation Complexes and Co-transcriptional CTD Phosphorylation-dependent RNA Capping
  • 10:26Conclusion

Summary

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Here, we describe the assembly of RNA polymerase II (Pol II) elongation complexes requiring only short synthetic DNA and RNA oligonucleotides and purified Pol II. These complexes are useful for studying mechanisms underlying co-transcriptional processing of transcripts associated with the Pol II elongation complex.

Tags

Keywords: RNA Polymerase IIElongation ComplexCo-transcriptional RNA ProcessingImmobilizationMagnetic BeadsDNA OligoBiotinWash BufferAnnealingThermal CyclerRNA-DNA HybridRNA Polymerase II Buffer

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