Workshop Description
Advances in Genetic Code Expansion and bioorthogonal ligations have changed the landscape of protein studies by enabling site-specific conjugation to proteins in vitro and in vivo. Genetically encoding bioorthogonal ligations is inherently a fusion of synthetic biology and chemical biology. Success requires 1) the addition of all the necessary translational components to an organism that allow it to efficiently produce a protein with new chemical functionality site-specifically encoded, 2) optimization of scientific studies of the chemically modified protein in vivo or in vitro, and 3) optimization of the labeling reaction to the protein of interest while minimizing any background reactions. Overall, success with genetically encoding bioorthogonal ligations relies on proficiency in both synthetic biology and chemical biology.
This intensive laboratory and lecture course will provide participants with the theoretical and practical knowledge to utilize existing and emerging genetic code expansion technology in biorthogonal ligations. The laboratory component will focus on using bioorthogonal ligation tools in E. coli and mammalian systems. Participants will learn strategies for appropriate controls and troubleshooting for all bioorthogonal GCE experiments, but in lab we will focus on Alkene and tetrazine inverse-demand Diels-Alder chemistry.
The laboratory component will focus on two common needs in the genetically encoding bioorthogonal ligation field: 1) incorporation of reactive non-canonical amino acid into your protein of interest and control proteins 2) labeling and reacting your protein with fluorescent dyes. As part of the laboratory component of the course, participants will be given the opportunity to bring their research gene of interest to the workshop to test their expression and incorporation along with optimized controls.
The lecture component will focus on how new genetic code expansion components are generated and attributes of orthogonality, efficiency, fidelity and permissivity are defined, measured and are conditionally dependent. The major applications now afforded by encoding bioorthogonal reactive non-canonical amino acids will be discussed with a focus on the limits to each technology and future applications.