Engineered Adenosine Deaminases for High-Efficiency Base Editing
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Solution Overview
Problem
Current adenine base editors (ABEs) such as ABE7.10 and miniABEmax are inefficient and take a long time to complete their reaction, while efficient adenosine deaminases like TadA-8e are rare, limiting their commercial applications.
Innovation Solution
Development of adenosine deaminases with specific amino acid substitutions at sites such as W23, Y36, P48, H51, L84, A106, D108, V109, K110, T111, D119, G122, H123, S146, F149, R152, H156, K157, E168, and E169, which enhance the efficiency of base editing processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If current ABE systems (ABE7.10, miniABEmax) are used, then base editing can be performed, but the reaction takes a long time and functions at low efficiency
Solution Approach 1:
The patent applies parameter changes by modifying amino acid residues at specific positions (K110, T111, D119, G122, H123) in the adenosine deaminase domain. These parameter changes in the protein structure directly improve the catalytic efficiency and reaction speed of the base editor, resolving the contradiction between editing capability and reaction efficiency.
Solution Approach 2:
The invention applies local quality by making targeted amino acid substitutions at specific positions within the adenosine deaminase domain rather than modifying the entire protein. This localized modification approach improves base editing efficiency while maintaining the overall structure and function of the base editor system.
2Productivity
If adenosine deaminases as efficient as TadA-8e are used, then base editing efficiency is improved, but such enzymes are rare and limit commercial uses
Solution Approach 1:
The patent creates a universal base editor system by optimizing the adenosine deaminase domain to achieve high efficiency comparable to rare TadA-8e, while making it compatible with standard CRISPR-Cas9 components. This makes the system broadly applicable for commercial use rather than limited to specialized applications.
Solution Approach 2:
By changing the amino acid parameters at key positions in the adenosine deaminase, the invention produces an enzyme with improved catalytic properties that is both highly efficient and commercially viable, overcoming the rarity limitation of TadA-8e.
3Productivity
If amino acid substitutions are made at multiple sites (W23, Y36, P48, H51, L84, A106, D108, V109, K110, T111, D119, G122, H123, S146, F149, R152, H156, K157, E168, E169), then base editing efficiency is enhanced, but the complexity of protein engineering increases
Solution Approach 1:
The patent applies local quality by focusing amino acid substitutions on specific functional regions of the adenosine deaminase domain. Rather than random modifications throughout the protein, the changes are concentrated at positions that directly impact catalytic efficiency, thereby enhancing base editing while managing engineering complexity.
Solution Approach 2:
The invention systematically changes amino acid parameters at identified key positions to optimize base editing efficiency. This structured parameter change approach based on structure-function relationships makes the complex protein engineering process more controllable and predictable.
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
The modified adenosine deaminases significantly improve the efficiency and speed of base editing, making them more suitable for commercial and medical applications.
Implementation Method 1
The adenosine deaminase, when the fusion protein targets the genomic DNA under the guidance of the guiding RNA (such as sgRNA), binds the ssDNA and converts adenine (A) to inosine (I)
Data Source
AI summary
Disclosed are adenosine deaminases, base editors comprising the adenosine deaminases and complexes comprising the base editors. The adenosine deaminases and the base editors exhibited superior adenine editing effects and achieved A·T base pair to G·C base pair transformation at DNA level.