CRISPR/Cas Compositions for Mutant SERPINA1 Editing
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Solution Overview
Problem
Alpha-1 antitrypsin deficiency (AATD) leads to both liver and lung pathology due to mutations in the SERPINA1 gene, causing insufficient and improperly formed alpha-1 antitrypsin (AAT), resulting in uncontrolled neutrophil elastase activity and protein aggregation in hepatocytes.
Innovation Solution
The CRISPR/Cas system is employed to deliver a composition comprising a guide RNA and an RNA-guided DNA binding agent to induce double-strand breaks in the SERPINA1 gene, reducing or preventing the production of mutant AAT forms associated with AATD.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-generated harmful factors
If CRISPR/Cas system is used to knock out SERPINA1 gene, then production of mutant AAT is eliminated, but complete elimination of AAT production may worsen lung protection function
Solution Approach 1:
The patent applies local quality by using allele-specific guide RNAs that target only the mutant SERPINA1 allele while leaving the wild-type allele intact. This selective knockout approach eliminates harmful mutant AAT production in the liver while preserving wild-type AAT synthesis for lung protection, thereby resolving the contradiction between eliminating liver pathology and maintaining lung function.
2Manufacturing precision
If CRISPR/Cas system delivers composition to hepatocytes, then SERPINA1 gene editing is achieved, but delivery efficiency and specificity to liver cells must be optimized
Solution Approach 1:
The patent uses lipid nanoparticles as intermediary delivery vehicles that encapsulate and protect the CRISPR/Cas9 components (guide RNA and Cas9 protein or mRNA) during transport to hepatocytes. These lipid nanoparticles facilitate efficient cellular uptake while protecting the sensitive nucleic acid components from degradation, thereby achieving precise gene editing without requiring overly complex delivery mechanisms.
3Productivity
If guide RNA sequences are optimized for high binding affinity, then editing efficiency increases, but off-target effects may increase
Solution Approach 1:
The patent designs guide RNAs with highly specific sequences that match only the mutant SERPINA1 allele, using precise nucleotide matching to ensure high binding affinity for the target while avoiding off-target effects. The guide RNA sequences are carefully selected to distinguish between mutant and wild-type alleles, achieving high editing efficiency with minimal off-target activity through localized sequence optimization.
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 effectively reduces AAT levels in the liver and serum, preventing protein accumulation and improving liver and lung symptoms in AATD patients by up to 99%.
Implementation Method 1
The CRISPR/Cas system is employed to deliver a composition comprising a guide RNA and an RNA-guided DNA binding agent to induce double-strand breaks in the SERPINA1 gene
Data Source
AI summary
Compositions and methods for introducing double-stranded breaks within the SERPINA1 gene are provided. Compositions and methods for reducing and eliminating mutant forms of α1-antitrypsin (AAT), such as seen in subjects having α1-antitrypsin deficiency (AATD), are provided.