AsCRISPR

AsCRISPR designs allele-specific guide RNAs (gRNAs) that discriminate mutant and wild-type alleles to enable selective disruption of disease-causing single-nucleotide variants and short indels for precision genome editing and allele-specific gene therapy.

Key Features:

• Allele-Specific Targeting: Designs gRNAs that discriminate single-base mutations and short insertion-deletion (indel) variants to enable selective disruption of mutant alleles while preserving wild-type alleles.
• Database Integration: Processes user-provided query sequences and integrates dbSNP, ClinVar, and OMIM data to analyze heterozygous SNPs and dominant single nucleotide variants (SNVs).
• CRISPR Nuclease Options: Supports multiple CRISPR nucleases and engineered variants, including Cas12b and CasX.
• Efficiency and Specificity Analysis: Evaluates on-target efficiencies and specificities of gRNA candidates and reports potential off-target effects and allele-specific restriction enzyme sites that may be disrupted by edits.
• Databases for gRNA Candidates: Generates a Dominant Database of candidate discriminating gRNAs targeting alternative alleles at dominant SNV sites and a manually curated Validated Database of experimentally validated discriminating gRNAs.

Scientific Applications:

• Allele-specific gene therapy: Enables selective disruption of mutant alleles or disease-causing SNPs while preserving normal alleles for therapeutic applications.
• Inherited diseases: Supports design of discriminating gRNAs for dominantly inherited conditions.
• Mutation-driven cancers: Facilitates targeting of cancer-associated driver mutations with allele-specific gRNAs.
• Genome imprinting: Enables allele-specific manipulation of imprinted loci.
• Haploinsufficiency: Allows allele-specific editing strategies relevant to haploinsufficiency contexts.
• Genome loci imaging: Supports design of allele-specific guides for locus-specific imaging.
• Immunocompatible manipulations: Supports allele-specific modifications for immunocompatible interventions.

Methodology:

Processes user-provided query sequences and integrates dbSNP, ClinVar, and OMIM to analyze heterozygous SNPs and dominant SNVs; designs allele-discriminating gRNAs for single-base mutations and short indels and supports multiple CRISPR nucleases including Cas12b and CasX; evaluates on-target efficiencies and specificities, identifies potential off-targets and allele-specific restriction enzyme sites disrupted by edits; and generates Dominant and manually curated Validated databases of discriminating gRNAs.