The most well characterized cell-free system for in vitro transcription of eukaryotic genes is derived from HeLa cell nuclei. HeLa nuclear extracts can support accurate transcription initiation by RNA polymerase II and exhibit both basal and regulated patterns of RNA polymerase transcription. The nuclear extract is also a source of transcription factors, DNA-binding proteins and the enzymatic machinery involved in RNA processing.
The HeLa Nuclear Extract included in the HeLaScribe® Nuclear Extract in vitro Transcription System is prepared by a modification of the method of Dignam et al. Extracts prepared by this method have been shown to allow transcription from the human transferrin gene promoter and the adenovirus 2 major late promoter.
The system includes all the necessary components for in vitro transcription as well as a positive control template (CMV immediate early promoter DNA), and is performance-tested with the cytomegalovirus immediate early gene (CMV) promoter. HeLaScribe® Nuclear Extract is also available separately.
Applications of the HeLaScribe® Nuclear Extract in vitro Transcription System include:
- Analyzing basal and transcription factor-regulated transcription.
- Assessing Promoter strength.
- Mapping mRNA start sites.
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