Expanding the Genetic Code of Mus Musculus
Professor Hee-Sung Park of the Department of Chemistry, who garnered attention for his novel strategy of installing authentic post-translational modifications into recombinant proteins, expanded his research portfolio to another level. Professor Park’s team was the first to report the generation of a mouse strain with an expanded genetic code, allowing site-specific incorporation of unnatural amino acids.
Professor Park published the research on the new chemical biology method for achieving selective chemical modifications in proteins in Science last September. The research team, this time in collaboration with Professor Chan Bae Park of the Department of Physiology at the Ajou University School of Medicine, demonstrated temporal and spatial control of protein acetylation in various organs of the transgenic mouse using a recombinant green fluorescent protein as a model protein. This research was published in the online edition of Nature Communications on February 21.
This approach enables the rapid onset of position-specific acetylation of a target protein at any developmental stage, facilitating temporal and spatial control of protein acetylation in various organs of the transgenic mouse. Such temporal and spatial control of protein acetylation will be of prime importance for investigating many essential biological processes and human diseases at the tissue and organism level.
Almost all human proteins, the products of about 25,000 genes, are known to undergo various post-translational modifications during and after synthesis. Post-translation modifications regulate the function of cellular proteins, playing a key role in many essential processes such as delivering signals and body growth. However, the unusual protein modifications, aroused from genetic and/or environmental factors, trigger severe diseases including cancer, dementia, and diabetes.
The team inserted transgenes into the mouse genome to allocate the site-specific addition of unnatural amino acids. The researchers inserted a modified version of lysine into the house mice, which allowed for the control of the acetylation. They used recombinant green fluorescent proteins from transgenic house mice as models for control of the acetylation.
The team was also able to regulate the acetylation of specific temporal and spatial frames in the mice, restraining the abnormality in proteins to certain organs such as the liver and kidneys. The research team said the strategy will provide a powerful tool for systematic in vivo study of cellular proteins in the most commonly used mammalian model organisms for human physiology and disease. Professor Park said, “This method can be easily extended to generate a wide range of custom-made transgenic mouse strains for further investigating diverse proteins of interest.” He added, “This method can be further extended to generate a wide range of custom-made transgenic mouse strains, opening a new paradigm for investigating anti-cancer and cerebral disease treatments.
This work was supported by grants from KAIST Systems Healthcare and the Medicinal Bioconvergence Research Center and the Intelligent Synthetic Biology Center of the Global Frontier Project funded by the Ministry of Science, ICT & Future Planning and the Ministry of Food and Drug Safety.
(Figure:Temporal and spatial control of in vivo protein acetylation)
(a) Temporal expression of acetylated GFPuv in the AcK-GFPamber mouse. The expression of GFPuv in skeletal muscle, liver, and lung tissues was detected only in the AcK-injected mouse. Scale bar, 200 µm. (b) Western blotting of anti-FLAG-immunoprecipitated proteins from tissues of the AcK-GFPamber mouse. Acetylated GFPuv was produced after AcK injection. (c) Spatial expression of acetylated GFPuv in the AcK-GFPamber mouse. Acetylated GFPuv was observed only in skeletal muscle when AcK was directly delivered to the tissues. Sacle bar, 200 µm.