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Efficient expression optimization of proteins and pathways using the RBS calculator

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Published on May 5, 2015

Maximizing a heterologous protein's titer can be a balancing act between under- and over-expression. Too much: the protein aggregates. Too little: scale-up is not feasible. This challenge becomes enormous when multiple proteins are involved, for example, when expressing a multi-subunit therapeutic or engineering a multi-enzyme metabolic pathway. It's costly and honestly quite boring to screen combinatorial libraries.

To solve these challenges, this webinar introduces the computational design of synthetic DNA to systematically optimize expression levels in one- or multi-protein genetic systems, while performing the fewest number of experimental measurements. We introduce the physics behind the RBS Calculator, a thermodynamic model that predicts a mRNA's translation initiation rate using only its sequence. We then show how to use the RBS Library Calculator to design small RBS libraries that search large expression spaces. Instead of screening 10^8 variants, our computational design approach requires only 10 to 100 characterized constructs. We present several case studies showing how to optimize protein titers and engineer multi-enzyme metabolic pathways.

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