Chemically Modified Nucleic Acids Modulate Cpf1 Nuclease Activity
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods lack effective ways to regulate the CRISPR-Cpf1 system, particularly for upregulation, downregulation, and complete inactivation, which is crucial for therapeutic applications due to potential side effects.
Innovation Solution
Design and synthesis of novel nucleic acid molecules, including chemically modified nucleotides and chimeric DNA/RNA guide molecules, that can act as adjustable switches to modulate the cleavage activity of Cpf1 in the CRISPR genome editing system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the CRISPR-Cpf1 system is used for genome editing, then gene editing efficiency is improved, but potential side effects increase
Solution Approach 1:
The patent introduces chemically modified nucleic acid molecules as intermediary substances that bind to the guide RNA and modulate Cpf1 activity. These molecules act as mediators between the guide RNA and the nuclease, enabling controlled regulation of gene editing efficiency and providing a mechanism to mitigate side effects through selective inhibition or enhancement of nuclease activity.
Solution Approach 2:
The patent employs chemically modified nucleotides (such as phosphorothioate modifications) that change the chemical parameters of the nucleic acid molecules. These parameter changes alter the binding affinity and interaction dynamics between the guide RNA and Cpf1, thereby enabling precise control over nuclease activity to balance editing efficiency with safety.
2Object-affected harmful factors
If chemically modified nucleic acid molecules are introduced to regulate Cpf1 activity, then side effects are mitigated, but system complexity increases
Solution Approach 1:
The patent utilizes short, chemically modified nucleic acid molecules (oligonucleotides) that can be easily synthesized and degraded. These disposable-like molecules provide temporary, controllable regulation of Cpf1 activity without requiring complex, long-lasting systems. Their short lifespan allows for rapid adjustment and elimination of regulatory function when needed.
Solution Approach 2:
By modifying the chemical parameters of the nucleic acid molecules (such as incorporating phosphorothioate linkages), the patent achieves effective regulation with relatively simple molecular structures. These parameter changes enable potent inhibitory or activating effects while maintaining acceptable structural simplicity, avoiding the need for highly complex regulatory systems.
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
These nucleic acid molecules effectively inhibit or enhance the nuclease activity of the CRISPR-Cpf1 system in a controlled manner, providing a mechanism to manage gene editing efficiency and mitigate side effects.
Implementation Method 1
the nucleic acid is complementary to the guide RNA
Implementation Method 2
the nucleic acid is complementary to the guide RNA
Data Source
AI summary
The present disclosure generally relates to genome editing systems and methods and compounds and compositions for use in Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) genome editing systems. Disclosed herein are modified nucleic acids that modulate the activity of genome editing.