Identification of anti-repressor elements that confer high and stable protein production in mammalian cells

The expression of transgenic proteins is often low and unstable over time, a problem that may be due to integration of the transgene in repressed chromatin. We developed a screening technology to identify genetic elements that efficiently counteract chromatin-associated repression. When these elemen...

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Bibliographic Details
Published in:Nature biotechnology 2003-05, Vol.21 (5), p.553-558
Main Authors: Otte, Arie P, Kwaks, Ted H.J, Barnett, Phil, Hemrika, Wieger, Siersma, Tjalling, Sewalt, Richard G.A.B, Satijn, David P.E, Brons, Janynke F, van Blokland, Rik, Kwakman, Paul, Kruckeberg, Arle L, Kelder, Angèle
Format: Article
Language:English
Subjects:
Agriculture
Animals
Bioinformatics
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
CHO Cells
Chromatin - genetics
Chromatin - metabolism
Cricetinae
Cricetulus
Deoxyribonucleic acid
DNA
Fundamental and applied biological sciences. Psychology
Gene Expression Profiling - methods
Gene Expression Regulation - genetics
Humans
Life Sciences
Mammals
Molecular Sequence Data
Promoter Regions, Genetic
Protein Biosynthesis
Protein Engineering - methods
Proteins - genetics
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Repressor Proteins - genetics
Repressor Proteins - metabolism
Transgenes - genetics
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Summary:The expression of transgenic proteins is often low and unstable over time, a problem that may be due to integration of the transgene in repressed chromatin. We developed a screening technology to identify genetic elements that efficiently counteract chromatin-associated repression. When these elements were used to flank a transgene, we observed a substantial increase in the number of mammalian cell colonies that expressed the transgenic protein. Expression of the shielded transgene was, in a copy number–dependent fashion, substantially higher than the expression of unprotected transgenes. Also, protein production remained stable over an extended time period. The DNA elements are small, not exceeding 2,100 base pairs (bp), and they are highly conserved between human and mouse, at both the functional and sequence levels. Our results demonstrate the existence of a class of genetic elements that can readily be applied to more efficient transgenic protein production in mammalian cells.
ISSN:1087-0156
1546-1696
DOI:10.1038/nbt814