Transgenic Mouse Model for Site-Specific Protein Modification
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Solution Overview
Problem
Current methods fail to temporally and spatially control site-specific modification of target proteins in mice, limiting detailed in vivo studies of protein functions and their roles in human diseases.
Innovation Solution
A transgenic mouse model is developed by introducing a tRNA synthetase specific for an unnatural amino acid, a tRNA that recognizes this amino acid, and a gene encoding a target protein with an amber codon, allowing for temporal and spatial control of site-specific protein modification through the incorporation of unnatural amino acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the amber codon suppression technique is used to enable site-specific modification of proteins, then protein functionality can be expanded at the molecular and cellular level, but the technique has not been successfully extended to multiorgan animal mice
Solution Approach 1:
The invention divides the complex system into separate transgenic components: (1) a first transgene encoding an orthogonal aminoacyl-tRNA synthetase and tRNA for amber codon suppression, and (2) a second transgene encoding the target protein with amber codons at specific positions. This segmentation allows each component to be optimized independently and combined in mice, resolving the complexity barrier while maintaining protein functionality expansion capability.
2Reliability
If conventional methods are used for studying protein functions in mice, then general protein expression can be achieved, but temporal and spatial control of site-specific modification is not possible
Solution Approach 1:
The invention introduces an intermediary control mechanism using unnatural amino acids (e.g., acetyl-lysine analogs) that serve as mediators between the researcher and the target protein. By feeding or injecting these small molecule intermediaries, researchers can temporally and spatially control the incorporation of modified amino acids into target proteins at amber codon sites, enabling precise control while maintaining reliable in vivo study conditions.
3Adaptability or versatility
If site-specific protein modification is achieved without temporal and spatial control, then protein diversity can be expanded, but detailed in vivo studies of protein functions remain limited
Solution Approach 1:
The invention performs preliminary action by pre-installing amber codons at specific positions in the target protein gene sequence before expression in mice. This preliminary genetic modification creates predetermined sites where site-specific modification can occur, allowing researchers to later control when and where modifications happen by introducing unnatural amino acids, thereby preserving spatial and temporal information that would otherwise be lost.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables precise temporal and spatial control of protein expression and modification in mice, facilitating in vivo studies of protein functions and their roles in human diseases, including cancer and neurodegenerative disorders.
Implementation Method 1
a tRNA synthetase specific for an unnatural amino acid
Data Source
AI summary
The present invention relates to a mouse (Mus musculus) 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.