{
  "abstract": "**Background** Canavan disease is a rare autosomal recessive neurodegenerative leukodystrophy caused by pathogenic variants in the ASPA gene, for which effective molecularly targeted therapies remain limited. The NM_000049.4:c.691T>C (p.Tyr231His) variant introduces a pathogenic C-G base pair that is potentially amenable to correction using cytosine base editing (CBE). This study aimed to design and computationally evaluate a CBE-based CRISPR strategy for potential correction of this ASPA variant. **Methods** An integrated in silico workflow was used to characterize the ASPA p.Tyr231His variant and design a CBE-compatible sgRNA. Variant pathogenicity, conservation, and structural stability were assessed using multiple computational tools. Candidate sgRNAs were designed and evaluated for editing-window positioning, PAM compatibility, specificity, GC content, bystander editing, and secondary structure. Genome-wide off-target potential was further assessed using CRISPRitz against the GRCh38 reference genome. **Results** The ASPA p.Tyr231His variant was exceptionally rare (gnomAD allele frequency: 1.86 x 10^-6), highly conserved, and consistently predicted to be deleterious, with DynaMut2 indicating reduced protein stability (DDG = -2.04 kcal/mol). Of two candidate sgRNAs, one was selected based on optimal editing-window positioning, absence of predicted bystander editing, favourable GC content (45%), and specificity scores (MIT: 82; CFD: 93). RNAfold predicted minimal secondary-structure constraints (MFE: 0.00 kcal/mol). CRISPRitz identified 108 potential off-target sites, with none containing fewer than three mismatches, no bulge-associated sites, and only three predicted sites within exonic regions. **Conclusions** The integrated computational analysis identified a candidate sgRNA with favourable predicted editing characteristics and genome-wide specificity for potential CBE-mediated correction of the ASPA c.691T>C (p.Tyr231His) variant. These findings provide a rational framework for prioritizing this guide for experimental evaluation. Further cell-based and in vivo studies are required to establish editing efficiency, editing purity, bystander activity, off-target effects, and restoration of ASPA function before therapeutic applicability can be assessed.",
  "abstract_shown": true,
  "affiliations": [
    "Biotechnika Info Labs"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 2,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "A. Verma"
    },
    {
      "name": "E. Natrajan"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/fqq4ygkf4jdkv.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.24.761534",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.24.761534v1.full",
  "id": "fqq4ygkf4jdkv",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.24.761534v1",
  "language": null,
  "licence": "CC-BY-NC",
  "licence_label": "CC BY-NC",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "bioinformatics"
    ],
    [
      "openalex",
      "1312|CRISPR and Genetic Engineering"
    ]
  ],
  "openalex": "W7223960476",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.24.761534v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-24",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/fqq4ygkf4jdkv.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:29:48Z",
      "native_id": "10.64898/2026.10.24.761534",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.24.761534v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:bioinformatics",
      "subfield": "bioinformatics"
    }
  ],
  "title": "In Silico Design and Evaluation of CRISPR Cytosine Base Editing Strategy for Potential Correction of the ASPA Variant Associated with Canavan Disease",
  "title_text": "In Silico Design and Evaluation of CRISPR Cytosine Base Editing Strategy for Potential Correction of the ASPA Variant Associated with Canavan Disease",
  "type": "preprint",
  "updated_at": "2026-10-30T12:29:48Z",
  "url": "https://research.draconitic.com/paper/fqq4ygkf4jdkv",
  "venue": "bioRxiv",
  "versions": []
}