PT  - JOURNAL ARTICLE
AU  - Junetha Syed
AU  - Saravanan Palani
AU  - Scott T. Clarke
AU  - Zainab Asad
AU  - Andrew R. Bottrill
AU  - Alexandra M.E. Jones
AU  - Karuna Sampath
AU  - Mohan K Balasubramanian
TI  - Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine
AID  - 10.1101/631432
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 631432
4099  - http://biorxiv.org/content/early/2019/05/10/631432.short
4100  - http://biorxiv.org/content/early/2019/05/10/631432.full
AB  - Site-specific incorporation of un-natural amino acids (UNAA) is a powerful approach to engineer and understand protein function [1-4]. Site-specific incorporation of UNAAs is achieved through repurposing the amber codon (UAG) as a sense codon for the UNAA, a tRNACUA that base pairs with an UAG codon in the mRNA and an orthogonal amino-acyl tRNA synthetase (aaRS) that charges the tRNACUA with the UNAA [5, 6]. Here, we report expansion of the zebrafish genetic code to incorporate the UNAAs, Azido-lysine (AzK), bicyclononyne-lysine (BCNK), and Diazirine-lysine (AbK) into green fluorescent protein (GFP) and Glutathione-S-transferase (GST). We also present proteomic evidence for UNAA incorporation into GFP. Our work sets the stage for the use of UNAA mutagenesis to investigate and engineer protein function in zebrafish.