Modular TCR Library System for Rapid Receptor Assembly
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Solution Overview
Problem
Current methods for generating and characterizing T cell receptors (TCRs) are economically inefficient and inflexible, requiring synthesis of multiple DNA sequences for each candidate TCR, and do not allow for easy alteration of C region improvements or exchange of TCR sub-domains, limiting the ability to rapidly reconstruct TCR sequences and evaluate different configurations.
Innovation Solution
A modular TCR library system comprising 45 TCR α chain and 47 TCR β chain constructs, each with variable, constant, and linker sequences, integrated into ivtRNA, retroviral, or lentiviral backbone vectors, enabling easy exchange of segments and flexible expression in T cells for therapeutic and immunization applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If DNA synthesis of full length TCR constructs is used, then complete TCR sequences can be obtained, but the process becomes non-economical and time-consuming when a wide variety of candidate TCRs must be functionally characterized
Solution Approach 1:
The TCR construct is divided into separate modular segments: variable region (V), constant region (C), and CDR3 region. Each segment can be independently synthesized and assembled, allowing rapid reconstruction of different TCR sequences by exchanging only the variable and CDR3 segments while reusing constant regions, thereby reducing synthesis costs and increasing throughput
Solution Approach 2:
The constant regions (AC and BC segments) are pre-synthesized and prepared in advance as reusable building blocks. This preliminary preparation eliminates the need to synthesize complete TCR sequences from scratch for each candidate, enabling rapid assembly and functional characterization of multiple TCR variants
2Manufacturing precision
If DNA synthesis of full length TCR constructs is used, then TCR sequences can be obtained, but the process lacks flexibility for retroactive alterations of C region improvements or exchange of TCR sub-domains
Solution Approach 1:
The TCR construct is segmented into distinct modular components (V region, C region, CDR3 region) that can be independently manipulated. This segmentation enables flexible recombination and exchange of specific TCR sub-domains without affecting other regions, allowing retroactive improvements and configuration changes
Solution Approach 2:
The constant region segments are designed as universal building blocks that can be reused across multiple TCR constructs. This multi-functionality allows the same constant region to be paired with different variable and CDR3 regions, enabling flexible reconstruction and evaluation of various TCR configurations
Data Source
Figure 1A~1B
Figure 2
Figure 3a~3
AI summary
The invention relates to a library for the expression of all functional TCR types comprising 45 TCR constructs each encoding one of the 45 different TCR α chains and 47 TCR constructs each encoding one of the 47 different TCR β chains, wherein each of the 45 TCR constructs encoding one of 45 different TCR α chain comprises the following building blocks one of the variable AV segments AVseg1 to AVseg45, and a constant AC segment, and wherein each of the 47 TCR constructs encoding one of 47 different TCR β chains comprises one of the variable BV segments BVseg1 to BVseg47, and a constant BC segment.