CpG-island fragments from the HNRPA2B1/CBX3 genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells.
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.


CONCLUSIONS
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.
Williams
Steven
S
ML Laboratories PLC-Research Division, MED IC4, Keele University Science and Business Park, Keele, Staffordshire, ST5 5SP, UK. Steve.Williams@mlresearch.co.uk
Mustoe
Tracey
T
Mulcahy
Tony
T
Griffiths
Mark
M
Simpson
David
D
Antoniou
Michael
M
Irvine
Alistair
A
Mountain
Andrew
A
Crombie
Robert
R
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics*; D002874:Chromosome Mapping; D018899:CpG Islands; D006224:Cricetinae; D003587:Cytomegalovirus / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005434:Flow Cytometry; D015870:Gene Expression; D005786:Gene Expression Regulation; D020868:Gene Silencing; D017930:Genes, Reporter; D005821:Genetic Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000737:chemistry; D014158:Transcription, Genetic; D014162:Transfection; D019076:Transgenes
C453799:Cbx3 protein, mouse; D002868:Chromosomal Proteins, Non-Histone; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins