Current Synthetic and Systems Biology
Open Access

DNA recombination engineering for improved and more reliable protein expression from DNA vectors

International Conference on Synthetic Biology

September 28-29, 2015 Houston, USA

Nicolas Mermod

University of Lausanne, Switzerland

Posters-Accepted Abstracts: Curr Synthetic Sys Biol

Abstract :

Epigenetic regulatory DNA elements can be added to expression vectors to increase transgene genomic integration to dampen silencing and to provide overall a more reliable expression in cultured mammalian cells. Nevertheless, expression remains variable from one cell clone to the next. We have sequenced the genomes of CHO cell clones expressing recombinant proteins yielding information on the molecular mechanisms that allowed the genomic integration of the plasmid vectors. This provided an approach to transiently silence unwanted recombination pathways so as to favor a microhomology-based recombination mechanism that mediates integration into expression-permissive genomic loci. When combined to MAR epigenetic regulatory elements, this provides much more predictable and homogeneous levels of transgene expression in polyclonal cell populations expressing a gene of interest.

Biography :

Email: Nicolas.Mermod@unil.ch