SYNTHETIC GENE DESIGN STRATEGIES FOR IMPROVING HETEROLOGOUS PROTEIN EXPRESSION AND PRODUCTION IN MICROBIAL AND MAMMALIAN CELLS

Abstract

The design and synthesis of synthetic genes is a key technology to accomplish rational host cell engineering in biotechnology and synthetic biology applications. In the design of synthetic genes, codon optimization is a proven strategy that can enhance protein expression. To optimize the target gene for expression in a host, the codon usage in the gene is modified algorithmically based on the selected design criteria and a reference codon bias. Consequently, the choice of design criteria and reference codon bias are crucial decisions in ensuring success. This work studies how the selection of codon bias representing hosts and selected design criteria affect the final gene design. By leveraging on available –omics profiling data, appropriate reference codon biases were established for industrially important hosts. The use of codon context as the key design criterion was also demonstrated to be superior to other commonly employed design criteria.