CpG-island fragments from theHNRPA2B1/CBX3genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells
✍ Scribed by Steven Williams; Tracey Mustoe; Tony Mulcahy; Mark Griffiths; David Simpson; Michael Antoniou; Alistair Irvine; Andrew Mountain; Robert Crombie
- Book ID
- 104497569
- Publisher
- BioMed Central
- Year
- 2005
- Tongue
- English
- Weight
- 731 KB
- Volume
- 5
- Category
- Article
- ISSN
- 1472-6750
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✦ Synopsis
Background
The hCMV promoter is very commonly used for high level expression of transgenes in mammalian cells, but its utility is hindered by transcriptional silencing. Large genomic fragments incorporating the CpG island region of the HNRPA2B1 locus are resistant to transcriptional silencing.
Results
In this report we describe studies on the use of a novel series of vectors combining the HNRPA2B1 CpG island with the hCMV promoter for expression of transgenes in CHO-K1 cells. We show that the CpG island gives at least twenty-fold increases in the levels of EGFP and EPO observed in pools of transfectants, and that transgene expression levels remain high in such pools for more than 100 generations. These novel vectors also allow facile isolation of clonal CHO-K1 cell lines showing stable, high-level transgene expression.
Conclusion
Vectors incorporating the hnRPA2B1 CpG island give major benefits in transgene expression from the hCMV promoter, including substantial improvements in the level and stability of expression. The utility of these vectors for the improved production of recombinant proteins in CHO cells has been demonstrated.