HeLaScribe® Nuclear Extract in vitro Transcription System

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 for a variety 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 p...

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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 for a variety 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 also includes all of the necessary components for in vitro transcription as well as a positive control template (CMV immediate early promoter DNA).

Features - Benefits

• Peformance-Tested: Tested with cytomegalovirus immediate early gene (CMV) promoter.
• Convenient: Available as a complete transcription system or extract alone.
• Positive Control: System contains a CMV promoter-positive control template.

Applications

• Assay basal and transcription factor-regulated transcription.
• Assay promoter strength.
• Source of transcription factors, DNA-binding proteins and RNA processing components.
• Map mRNA start sites.

References

1. Dignam, J.D. et al. (1983) Nucl. Acids Res. 11, 1475–89.
2. Sawadogo, M. and Roeder, R.G. (1985) Proc. Natl. Acad. Sci. USA 82, 4394–8.
3. Briggs, M.R. et al. (1986) Science 234, 47–52.
4. Bagchi, M.K. et al. (1990) J. Biol. Chem. 265, 5129–34.
5. Carthew, R.W. et al. (1985) Cell 43, 439–48.
6. Kristie, T.M. et al. (1989) EMBO J. 8, 4229–38.
7. Sawadogo, M. et al. (1988) J. Biol. Chem. 263, 11985–93.
8. Krainer, A.R. et al. (1984) Cell 36, 993–1005.

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