Cas Type V RNP and Lipid Nanoparticle Delivery for Precise Gene Editing
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Solution Overview
Problem
Current CRISPR-Cas type V systems face challenges in achieving sufficient editing efficiency, precision, deliverability, and affordability for treating genetic disorders and complex diseases.
Innovation Solution
Development of Cas TypeV-based gene editing systems comprising a Type V polypeptide and a Type V guide RNA that form a complex to target and bind to genomic sequences, with optional accessory proteins for enhanced editing capabilities, delivered through various methods including viral and non-viral vectors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CRISPR-Cas type V systems are used for gene editing, then genome editing capability is provided, but editing efficiency and precision are insufficient
Solution Approach 1:
The system divides the guide RNA into two separate components: a crRNA (crISPR RNA) that binds to the target DNA sequence and a tracrRNA (trans-activating crRNA) that binds to the Cas protein. This segmentation allows each component to be optimized independently for its specific function, improving both the efficiency of target recognition and the precision of editing outcomes.
Solution Approach 2:
The patent creates a composite ribonucleoprotein complex by combining the Cas protein with the segmented guide RNAs (crRNA and tracrRNA). This composite structure integrates the enzymatic activity of the Cas protein with the sequence-specific targeting capability of the guide RNAs, achieving both high editing efficiency and precision through synergistic interaction of multiple components.
2Reliability
If CRISPR-Cas type V systems are used for gene editing, then genome editing capability is provided, but deliverability is insufficient
Solution Approach 1:
By segmenting the guide RNA into separate crRNA and tracrRNA components, the system enables independent optimization of delivery mechanisms for each component. This allows use of various delivery vectors (viral or non-viral) that can efficiently transport individual RNA molecules or their combinations, improving deliverability to target cells while maintaining editing capability.
3Reliability
If CRISPR-Cas type V systems are used for gene editing, then genome editing capability is provided, but cost-effectiveness is insufficient
Solution Approach 1:
The system uses synthetic, chemically synthesized guide RNAs (crRNA and tracrRNA) that can be produced cost-effectively through standard molecular biology techniques. These disposable RNA components replace the need for complex, expensive protein-based delivery systems or viral vectors, making the gene editing technology more cost-effective while maintaining full editing capability.
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 system provides improved editing efficiency, precision, and cost-effectiveness for gene editing in cells, tissues, and organisms, offering potential therapeutic benefits for genetic disorders.
Implementation Method 1
a Type V guide RNA which is capable of associating with a Type V polypeptide to form a complex such that the complex localizes to a target nucleic acid sequence and binds thereto
Implementation Method 2
the Type V polypeptide has a nuclease activity which results in the cutting of both strands of DNA
Data Source
AI summary
The present disclosure provides methods and compositions comprising Cas TypeV programmable nucleases and lipid nanoparticles capable of delivering the Cas TypeV programmable nucleases and genome editing systems comprising same.