Split Cpf1 Protein Fragments for Controlled Genome Editing
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Solution Overview
Problem
Current genome editing techniques lack a novel approach using the Cpf1 protein, which is essential for precise and efficient DNA cutting and editing.
Innovation Solution
Development of a split Cpf1 protein by dividing the Cpf1 protein into N-terminal and C-terminal fragments that can associate either spontaneously or upon light or drug induction, allowing for controlled genome editing through dimer formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the Cpf1 protein is divided into two separate fragments (N-terminal and C-terminal), then the genome editing technique becomes novel and controllable through dimer formation, but the complexity of the system increases due to requiring two separate polypeptides to associate
Solution Approach 1:
The Cpf1 protein is divided into two separate polypeptides (N-terminal fragment and C-terminal fragment) that can independently function but require association to achieve full nuclease activity. This segmentation enables controlled genome editing through regulated dimer formation while maintaining the ability to perform precise DNA cutting when activated
Solution Approach 2:
The two separate Cpf1 fragments are designed to associate through dimer formation, merging their functions to restore full nuclease activity. This merging mechanism provides controllable activation where the fragments remain separate until conditions trigger their association, thereby controlling the genome editing process
2Productivity
If the split Cpf1 protein fragments associate spontaneously, then the genome editing occurs efficiently without external induction, but the control over timing and location of editing is reduced
Solution Approach 1:
The association state of the split Cpf1 fragments is made dynamic rather than fixed. The fragments can transition between associated and dissociated states based on environmental conditions, allowing the system to adapt between spontaneous high-efficiency editing and controlled timed editing depending on operational needs
3Reliability
If the Cpf1 protein is used as a whole without splitting, then the DNA cutting activity is immediate and strong, but the ability to control the timing and spatial distribution of editing is lost
Solution Approach 1:
The Cpf1 protein is segmented into two functional fragments that retain individual stability but require association for full nuclease activity. This segmentation maintains reliable DNA cutting capability when fragments associate while enabling control mechanisms through regulated association/dissociation dynamics
Solution Approach 2:
The association interface between the two Cpf1 fragments acts as an intermediary control point. By regulating whether the fragments associate or remain separate, the system controls when and where full nuclease activity is expressed, thereby controlling the timing and spatial distribution of editing events
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 split Cpf1 protein enables efficient and precise genome editing with enhanced DNA cutting efficiency, offering both inducible and spontaneous association modes for targeted gene modification and expression control.
Implementation Method 1
the split Cpf1 protein is rearranged through induced association or spontaneous association
Data Source
AI summary
The present invention provides a set of two polypeptides of a split Cpf1 protein, wherein the two polypeptides are a N-terminal side fragment of a Cpf1 protein and a C-terminal side fragment of the Cpf1 protein.