CRISPR Screening Strategy for ASD Mouse Models
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Solution Overview
Problem
Current methods for genome editing, particularly with the CRISPR-Cas9 system, face challenges in efficiently delivering genetic modifications in vivo and ex vivo, especially in neuronal cells, which are sensitive and hard to access, limiting the study of genetic mutations associated with autism spectrum disorder (ASD) and other neurological conditions.
Innovation Solution
Development of a CRISPR-based screening strategy using vectors like AAV and lentiviral vectors to deliver guide RNAs and the Cas9 enzyme into neuronal cells, enabling multiplexed genome editing for modeling ASD and other neurological disorders, allowing for the rapid generation of mouse models with specific genetic mutations and the study of their phenotypic effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If CRISPR-Cas9 genome editing is applied in vivo and ex vivo, then precise genetic modifications can be achieved, but delivery efficiency is insufficient and accessible cell types are limited
Solution Approach 1:
Viral vectors (AAV, lentiviral vectors) are used as intermediary carriers to deliver CRISPR-Cas9 components into target cells. The patent employs these vectors to overcome the limitation of direct delivery methods, enabling efficient transduction of neuronal cells and other difficult-to-access cell types while maintaining precise genome editing capability
Solution Approach 2:
The CRISPR-Cas9 system is segmented into separate deliverable components (guide RNA and Cas9 enzyme) that can be individually packaged into viral vectors. This segmentation allows for optimized delivery of each component to target cells, improving overall delivery efficiency while preserving the precision of the genome editing mechanism
2Adaptability or versatility
If CRISPR-Cas9 is used to edit neuronal cells, then genetic mutations associated with neurological conditions can be studied, but neuronal cells are sensitive and hard to access
Solution Approach 1:
Viral vectors serve as intermediaries to bridge the gap between external delivery systems and protected neuronal cells. The patent utilizes AAV and lentiviral vectors that can efficiently transduce neuronal cells through their cell membranes, overcoming the accessibility barrier while enabling study of genetic mutations in these sensitive cells
Solution Approach 2:
The patent optimizes delivery parameters including vector choice, dosage, and administration route to achieve efficient delivery to neuronal cells. By adjusting these parameters, the system overcomes the difficulty of accessing neuronal cells while maintaining their sensitivity and enabling genetic mutation studies
Data Source
AI summary
Methods and compositions for inducing a plurality of mutations in transgenic Cas9 eukaryotes to model a neuronal disease or disorder. The invention further comprehends testing putative treatments with such models, e.g., testing putative chemical compounds that may be pharmaceutically relevant for treatment or gene therapy that may be relevant for treatment, or combinations thereof. The invention allows for the study of genetic diseases and putative treatments to better understand and alleviate a genetic disease or a condition, e.g., autism, autism-spectrum disease or disorder, obsessive-compulsive disorder, or psychiatric disorders.