CRISPR/Cas9 APP Gene Editing for Alzheimer's Aβ Reduction
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Solution Overview
Problem
Current methods fail to effectively reduce the production of toxic Aβ peptides, which are central to Alzheimer's disease pathology, limiting the prevention and treatment of the disease.
Innovation Solution
Gene editing using site-specific endonucleases and donor nucleic acids is employed to modify the APP gene, specifically reducing Aβ peptide production by introducing modifications such as the A673T substitution, which impairs BACE1 cleavage, thereby decreasing the levels of amyloidogenic peptides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current methods are used to reduce Aβ peptide production, then the existing treatment approaches are maintained, but the effectiveness in reducing toxic Aβ peptides is insufficient
Solution Approach 1:
The invention changes the molecular parameters of the APP gene by introducing specific nucleotide substitutions (such as A673T) that alter the amino acid sequence, thereby changing the cleavage sites for BACE1 and reducing Aβ peptide production. This parameter change at the genetic level directly addresses the insufficiency of current methods in reducing toxic Aβ peptides.
Solution Approach 2:
The invention replaces conventional pharmacological or mechanical treatment approaches with a molecular biology-based gene editing system. By using site-specific endonucleases and donor nucleic acids to directly modify the APP gene, the method substitutes traditional therapeutic mechanisms with a precise genetic intervention approach, achieving more effective Aβ reduction.
2Reliability
If gene editing methods are implemented to modify the APP gene, then Aβ peptide production is reduced, but the complexity of the treatment approach increases
Solution Approach 1:
The gene editing system is segmented into distinct functional components: guide RNA molecules that target specific APP gene sequences, site-specific endonucleases that create precise cuts, and donor nucleic acids that provide the desired genetic modifications. This segmentation allows each component to be optimized independently while working together to achieve the protective effect against Alzheimer's disease.
Solution Approach 2:
The invention uses guide RNA as an intermediary that bridges the target APP gene sequence and the site-specific endonuclease. The guide RNA contains sequences complementary to the target gene region, enabling precise localization of the editing machinery without requiring direct protein-DNA interaction, thereby simplifying the overall system architecture.
3Quantity of substance
If modifications are introduced to impair BACE1 cleavage, then amyloidogenic peptide production decreases, but the precision required for gene modification increases
Solution Approach 1:
The invention applies local quality changes by introducing specific nucleotide substitutions at precise locations within the APP gene (such as position 673) that locally alter the protein structure and cleavage susceptibility. This localized modification approach allows impairment of BACE1 cleavage without affecting other functional regions of the APP protein, thereby decreasing amyloidogenic peptide production with high precision.
Solution Approach 2:
The donor nucleic acids are designed in advance with the desired genetic modifications already incorporated, such as the A673T substitution that impairs BACE1 cleavage. This preliminary preparation of corrected gene sequences allows for precise and accurate introduction of the desired changes during the gene editing process, reducing the precision burden during actual modification.
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
This approach significantly reduces Aβ peptide production, providing a protective effect against Alzheimer's disease and age-related cognitive decline, as demonstrated by the increased likelihood of reaching old age without AD and reduced beta-amyloid pathology.
Implementation Method 1
A role of Amyloid β (Aβ) peptide aggregation and deposition in AD pathogenesis is widely accepted. Toxic peptides are produced by the metabolic processing of Amyloid β Precursor Protein (APP).
Implementation Method 2
The present application is a National Stage Application of PCT Application No. PCT/CA2015/050411 filed on May 8, 2015 and published in English under PCT Article 21(2), which claims the benefit of U.S. Provisional Application Ser. No. 61/991,054 filed on May 9, 2014.
Implementation Method 3
Gene editing using site-specific endonucleases and donor nucleic acids is employed to modify the APP gene, specifically reducing Aβ peptide production by introducing modifications such as the A673T substitution
Implementation Method 4
The aspartyl protease β-site APP cleaving enzyme 1 (BACE1) was originally identified over a decade ago (6, 7). It cleaves APP mostly at a unique site.
Data Source
AI summary
Methods and products are described related to use of the CRISPR/Cas9 system to introduce a modification into an APP gene, such as guide RNAs and recombinant proteins, for decreasing amyloid beta peptide produced by a cell. Also described are uses of such methods and products for the treatment of Alzheimer's disease and/or age-related cognitive decline in a cell from a subject in need thereof.