Cas Effector Domains for Programmable Gene Activation and Repression
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Solution Overview
Problem
Current synthetic transcription factors lack the full spectrum of regulatory mechanisms to effectively program gene expression in mammalian cells, limiting their potency and specificity.
Innovation Solution
Development of Cas effector domains, including polypeptides and antibodies, that can form fusion proteins with Cas proteins to modulate gene expression by targeting specific genes, using a range of effector proteins such as MCRS1, OTUD7B, RelB, LDB1, and others, which can increase or decrease gene expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If currently described effector domains are used in synthetic transcription factors, then the system is relatively simple to construct, but the full spectrum of regulatory mechanisms cannot be programmed, limiting gene expression control capability
Solution Approach 1:
The patent applies universality by developing a platform where a single Cas protein framework can be combined with multiple different effector domains (transcription activators, repressors, epigenetic modifiers) to achieve diverse gene regulation functions. This multi-functional approach allows one core system to program the full spectrum of regulatory mechanisms including transcriptional activation, repression, and epigenetic modifications.
Solution Approach 2:
The patent employs composite materials by creating fusion proteins that combine Cas proteins with various effector domains. These composite structures integrate the DNA-targeting capability of Cas proteins with the regulatory functions of effector domains, enabling programmable gene expression control through combinations of different molecular components.
2Reliability
If more effector domains are added to broaden regulatory mechanisms, then gene activation and repression potency and specificity improve, but system complexity increases
Solution Approach 1:
The patent applies segmentation by dividing the gene regulation function into modular components: a DNA-targeting module (Cas protein with guide RNA) and separate functional modules (different effector domains). This segmentation allows selective combination of components based on specific regulatory needs, improving control precision without requiring all effector domains to be present simultaneously.
Solution Approach 2:
The patent uses universality by creating a universal Cas protein platform that can interface with multiple types of effector domains. This universal framework enables the system to achieve high reliability in gene expression control through selective assembly of appropriate effector domains for specific applications, rather than requiring a fixed complex structure.
Data Source
AI summary
Disclosed herein are effector domains. The effector domains may be used with, for example, Cas proteins and CRISPR-Cas systems. The effectors may be used in combination with a Cas protein to form a fusion protein. The effectors may also be used in combination with an antibody that binds to a peptide epitope, wherein the peptide epitope is fused to a Cas protein. The compositions and methods comprising the effectors may be used to modulate gene expression.