Yale University

Korea Advanced Institute of Science and Technology

Hee-Sung Park

Dieter Soll

Aerin Yang

Hee-Yoon Lee

Sura Ha

The present invention relates to a mouse () in which expression and site-specific modification of a target protein is temporally and spatially controlled, and a method for producing the same and the use thereof, and more particularly to a transgenic mouse in which expression of a target protein having a modification attached to a specific position is temporally and spatially controlled as a result of incorporation of an unnatural amino acid. In the mouse according to the present invention, in which site-specific modification of a target protein is temporally and spatially controllable, expression of the target protein having the site-specific modification attached thereto is controllable depending on the timing and/or position of introduction of an unnatural amino acid. Thus, the mouse according to the present invention is useful for studies on the in vivo functions of cellular proteins, various human diseases including cancers and neurodegenerative disorders, new drug discovery, and the like.

Transgenic mouse capable of spatial and temporal control of expression and site-specific modification of target protein, production method and uses thereof

A method for producing a site-specifically modified protein based on new carbon-carbon bond formation is disclosed, including the following three steps (marking, activation, and coupling steps): (a) marking of the modification site by incorporating a specific amino acid into a selected position of a target protein; (b) activation of the marked site; and (c) coupling of various post-translational modification (PTM) moieties or other chemical groups onto the activated site to obtain a site-specifically modified protein. The method for producing a site-specifically modified protein can incorporate desired diverse chemical groups including post-translational modification (PTM) moieties into a designated site in a target protein through a new carbon-carbon bond. Furthermore, the modified protein having a site-specific PTM exhibits the same chemical and functional properties as that of a target protein present in cells. Thus, the present invention is useful for studies of cellular proteins, human diseases including cancers and neurodegenerative diseases, and new drug discovery.

Method for preparing site-specifically modified protein based on novel carbon-carbon bond formation

Nucleic acids encoding mutant elongation factor proteins (EF-Sep), phosphoseryl-tRNA synthetase (SepRS), and phosphoseryl-tRNA (tRNA) and methods of use in site specific incorporation of phosphoserine into a protein or polypeptide are described. Mutant EF-Sep proteins are disclosed that bind Sep-tRNAand protect Sep-tRNAfrom deacylation. In a preferred embodiment the nucleic acids are on vectors and are expressed in cells such as bacterial cells, archeaebacterial cells, and eukaryotic cells. Proteins or polypeptides containing phosphoserine produced by the methods described herein can be used for a variety of applications such as research, antibody production, protein array manufacture and development of cell-based screens for new drug discovery.

Site-specific incorporation of phosphoserine into proteins in

Nucleic acids encoding mutant elongation factor proteins (EF-Sep), phosphoseryl-tRNA synthetase (SepRS), and phosphoseryl-tRNA (tRNA) and methods of use in site specific incorporation of phosphoserine into a protein or polypeptide are described. Typically, SepRS preferentially aminoacylates tRNAwith O-phosphoserine and the tRNArecognizes at least one codon such as a stop codon. Due to the negative charge of the phosphoserine, Sept-tRNAdoes not bind elongation factor Tu (EF-Tu). However, mutant EF-Sep proteins are disclosed that bind Sep-tRNAand protect Sep-tRNAfrom deacylation. In a preferred embodiment the nucleic acids are on vectors and are expressed in cells such as bacterial cells, archeaebacterial cells, and eukaryotic cells. Proteins or polypeptides containing phosphoserine produced by the methods described herein can be used for a variety of applications such as research, antibody production, protein array manufacture and development of cell-based screens for new drug discovery.

Growing community of inventors

Daejeon, South Korea

Hee-Sung Park