|ZFIN ID: ZDB-PUB-190528-23|
Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine
Syed, J., Palani, S., Clarke, S.T., Asad, Z., Bottrill, A.R., Jones, A.M.E., Sampath, K., Balasubramanian, M.K.
|Source:||International Journal of Molecular Sciences 20(10): (Journal)|
|Registered Authors:||Clarke, Scott, Sampath, Karuna|
|Keywords:||click chemistry, crosslinking, genetic code expansion, protein engineering, proteomics, unnatural amino acid, zebrafish|
|PubMed:||31130675 Full text @ Int. J. Mol. Sci.|
Syed, J., Palani, S., Clarke, S.T., Asad, Z., Bottrill, A.R., Jones, A.M.E., Sampath, K., Balasubramanian, M.K. (2019) Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine. International Journal of Molecular Sciences. 20(10):.
ABSTRACTSite-specific incorporation of un-natural amino acids (UNAA) is a powerful approach to engineer and understand protein function. Site-specific incorporation of UNAAs is achieved through repurposing the amber codon (UAG) as a sense codon for the UNAA, using 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. Here, we report an 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 AzK, BCNK, and AbK introduction into proteins as a means to investigate and engineer their function in zebrafish.
ADDITIONAL INFORMATION No data available