Genome-wide prediction of matrix attachment regions that increase gene expression in mammalian cells.
Gene transfer in eukaryotic cells and organisms suffers from epigenetic effects that result in low or unstable transgene expression and high clonal variability. Use of epigenetic regulators such as matrix attachment regions (MARs) is a promising approach to alleviate such unwanted effects. Dissection of a known MAR allowed the identification of sequence motifs that mediate elevated transgene expression. Bioinformatics analysis implied that these motifs adopt a curved DNA structure that positions nucleosomes and binds specific transcription factors. From these observations, we computed putative MARs from the human genome. Cloning of several predicted MARs indicated that they are much more potent than the previously known element, boosting the expression of recombinant proteins from cultured cells as well as mediating high and sustained expression in mice. Thus we computationally identified potent epigenetic regulators, opening new strategies toward high and stable transgene expression for research, therapeutic production or gene-based therapies.
Institute of Biotechnology, University of Lausanne, and Center for Biotechnology of the University of Lausanne and Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Pierre-Alain
PA
Girod
Duc-Quang
DQ
Nguyen
David
D
Calabrese
Stefania
S
Puttini
Mélanie
M
Grandjean
Danielle
D
Martinet
Alexandre
A
Regamey
Damien
D
Saugy
Jacques S
JS
Beckmann
Philipp
P
Bucher
Nicolas
N
Mermod
D000818:Animals; D016466:CHO Cells; D002645:Chickens; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D015894:Genome, Human; D006801:Humans; D045170:Matrix Attachment Regions / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014162:Transfection; D019076:Transgenes
D011994:Recombinant Proteins