A Combinatorial Assembly Pipeline for Gene Expression in Mammalian Cells | AIChE

A Combinatorial Assembly Pipeline for Gene Expression in Mammalian Cells

Authors 

Stach, C. - Presenter, University of Minnesota, Twin Cities
Smanski, M. J., University of Minnesota, Twin Cities

Mammalian synthetic biology holds promise in developing cell based therapies, natural product synthesis, protein therapeutics, and defining the role of complex signaling networks in cell development. Progress towards these goals has been hampered by the limited availability of efficient and straightforward assembly pipelines in mammalian cells. While assembly pipelines exist for the generation of multi-gene constructs, they are limited in their utility. Currently, protein expression in mammalian cells leads to heterogeneous populations of the same protein due to differences in post-translational modifications. Here I present a combinatorial assembly pipeline developed for use in mammalian cells that can be used to identify genes important to expression of homogeneous protein populations. It is based on the use of Type IIS enzymes to assemble a series of defined parts such as promoters, terminators, and genes, into expression constructs. This system permits the expression of genes in a predictable manner and the ability to assemble and express genes from various pathways to examine their role within the cell.