Zinc Finger Peptide Synthesis via Fragment Segmentation
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Solution Overview
Problem
Current methods for introducing non-naturally occurring zinc fingers into cells or organisms are cumbersome and pose risks, particularly due to the use of recombinant nucleic acids, which complicates the creation of zinc finger libraries and raises concerns about foreign nucleic acid introduction.
Innovation Solution
The synthesis of zinc finger peptides and concatemers using specific peptide sequences, where the zinc finger domain is split into N-terminal and C-terminal fragments, allowing for reduced building blocks and facilitated sequence-specific binding, with options for labeling and membrane-penetrating peptides for enhanced detection and delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If recombinant nucleic acids are used to introduce zinc fingers into cells, then zinc finger expression can be achieved, but the process becomes cumbersome and risks associated with foreign nucleic acid introduction arise
Solution Approach 1:
The invention extracts the zinc finger peptide sequence from its traditional recombinant nucleic acid context and delivers it directly as a chemically synthesized peptide. This removes the foreign nucleic acid component while retaining the functional zinc finger domain, thereby eliminating safety risks associated with nucleic acid introduction while simplifying the manufacturing process for creating zinc finger libraries
Solution Approach 2:
The invention uses chemical synthesis as an intermediary method to produce zinc finger peptides directly without requiring recombinant nucleic acid transfection. This intermediary approach bypasses the need for DNA/RNA intermediates, enabling direct peptide delivery that is both safer and more efficient for creating diverse zinc finger libraries
2Adaptability or versatility
If recombinant DNA technology is used to create zinc finger libraries, then diverse zinc fingers can be generated, but the creation process becomes rather cumbersome
Solution Approach 1:
The invention segments the zinc finger domain into synthesizable peptide components that can be independently prepared and assembled. This segmentation allows for modular construction of diverse zinc finger sequences through chemical synthesis rather than complex recombinant DNA assembly, thereby maintaining sequence diversity while dramatically reducing process complexity
Solution Approach 2:
The invention changes the fundamental parameter of zinc finger production from genetic (recombinant DNA) to chemical (peptide synthesis). This parameter change enables direct synthesis of diverse zinc finger sequences with controlled amino acid composition, simplifying the library creation process while maintaining the ability to generate diverse sequences through systematic peptide assembly
Data Source
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AI summary
This invention relates to methods of producing zinc finger peptides and concatamers thereof as well as peptides of use in these methods. In particular, the invention relates to peptides consisting of the sequence set forth in any one of SEQ ID NOs: 1 to 3 and 6 to 8. Furthermore provided are methods of producing a zinc finger domain and concatemers thereof.