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types relevant to amyotrophic ","entities":{"genes":["HNRNPA2B1/SETX/TARDBP"],"diseases":["ALS"]},"model_system":"cell_line","_schema_version":1,"experiment_type":"falsification","primary_outcome":"Cell-type-specific base editing efficiency measured by single-cell RNA sequencing across motor neurons, astrocytes, and microglia, with comprehensive genome-wide off-target analysis.","methodology_notes":"**Phase 1: Cell Line Preparation and Editing (Weeks 1-2)**\n• Culture human iPSC-derived motor neurons (n=6 lines) and astrocytes (n=6 lines) representing ALS disease models\n• Prepare CRISPR-Cas9 editing constructs targeting ALS-associated genes (SOD1, C9orf72, TARDBP)\n• Perform electroporation-based gene editing with guide RNAs at 48-hour intervals\n• Maintain edited and control cell lines in parallel cultures with standard media conditions\n\n**Phase 2: Single-cell RNA-seq Analysis (Weeks 3-4)**\n• Harvest cells at 7, 14, and 21 days post-editing (n=10,000 cells per timepoint per line)\n• Process samples using 10x Genomics Chromium platform for scRNA-seq library preparation\n• Sequence libraries to achieve >50,000 reads per cell with >80% saturation\n• Perform bioinformatics analysis using Seurat pipeline for cell type identification and editing efficiency quantification\n• Calculate on-target editing rates using CRISPResso2 analysis of aligned reads\n\n**Phase 3: Genome-wide Off-target Detecti","replication_status":"replicated","extraction_metadata":{"backfill_at":"2026-04-16T01:00:16.911021","needs_review":true,"extraction_notes":"Backfilled from debate_extraction source (no PMID 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ALS"},{"target_artifact_id":"wiki-companies-als-pipeline","link_type":"related","strength":0.63,"evidence":"Reverse: Gene mention: ALS"},{"target_artifact_id":"wiki-therapeutics-rnai-therapies-neurodegeneration","link_type":"related","strength":0.63,"evidence":"Reverse: Gene mention: RNA"},{"target_artifact_id":"wiki-therapeutics-sod1-targeting-therapies","link_type":"related","strength":0.63,"evidence":"Reverse: Gene mention: SOD1"},{"target_artifact_id":"wiki-cell-types-microglia","link_type":"related","strength":0.54,"evidence":"Reverse: Title match: Microglia"},{"target_artifact_id":"wiki-diseases-als-ftd-spectrum","link_type":"related","strength":0.63,"evidence":"Reverse: Gene mention: ALS"},{"target_artifact_id":"wiki-cell-types-amyotrophic-lateral-sclerosis-frontotemporal-dementia-neurons","link_type":"related","strength":0.63,"evidence":"Reverse: Gene mention: 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FUS"},{"source_artifact_id":"wiki-mechanisms-neuroinflammation","link_type":"mentions","strength":0.6,"evidence":"Title match: neuroinflammation"},{"source_artifact_id":"wiki-mechanisms-als-combination-therapy-matrix","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-therapeutics-als-therapeutic-landscape","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-experiments-c9orf72-als-ftd-phenotype-mechanism","link_type":"mentions","strength":0.7,"evidence":"Gene mention: C9ORF72"},{"source_artifact_id":"wiki-technologies-als-communication-bci","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-als-fus-pathway","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-therapeutics-aav-vectors-neurodegenerative-gene-therapy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-technologies-rna-editing-therapeutics","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-proteins-fus-protein","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-entities-microglia","link_type":"mentions","strength":0.6,"evidence":"Title match: Microglia"},{"source_artifact_id":"wiki-mechanisms-sod1-superoxide-dismutase-als-causal-chain","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-mechanisms-ftd-fus-pathway","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-mechanisms-als-cure-roadmap","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-als-sod1-pathway","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-mechanisms-rna-g-quadruplexes-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-mechanisms-als-failed-approaches-analysis","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-therapeutics-fus-als-treatment","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-mechanisms-rna-stability","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-entities-heat-shock-proteins","link_type":"mentions","strength":0.6,"evidence":"Title match: Heat Shock Proteins"},{"source_artifact_id":"wiki-diseases-cns-vasculitis","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CNS"},{"source_artifact_id":"wiki-diseases-als","link_type":"mentions","strength":0.6,"evidence":"Title match: Amyotrophic Lateral Sclerosis 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RNA"},{"source_artifact_id":"wiki-diseases-als-ftd-parkinsonism-comparison","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-rna-dysregulation-ad","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-therapeutics-als-treatment","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-proteins-sod1","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-institutions-als-association","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-cns-drug-delivery-methods","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CNS"},{"source_artifact_id":"wiki-experiments-als-regional-onset-spread","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-genes-sod1","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-ideas-payload-aav-capsid-engineering-cns-therapy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-technologies-crispr-gene-editing-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-mechanisms-cbs-rna-metabolism-dysfunction","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-experiments-als-progression-rate-heterogeneity","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-genes-rbfox3","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-therapeutics-crispr-brain-delivery","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-experiments-aav-lrrk2-ind-enabling-studies","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-mechanisms-aav-gene-therapy-vectors-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-mechanisms-rna-therapeutics-investment-synthesis","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-mechanisms-als-progression-trajectories","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-therapeutics-crispr-cas9-gene-therapy-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-ideas-cns-nlrp3-inflammasome-inhibitor","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CNS"},{"source_artifact_id":"wiki-experiments-lrrk2-aav-ind-enabling-study","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-proteins-c9orf72-protein","link_type":"mentions","strength":0.7,"evidence":"Gene mention: C9ORF72"},{"source_artifact_id":"wiki-circuits","link_type":"mentions","strength":0.6,"evidence":"Title match: Circuits"},{"source_artifact_id":"wiki-cell-types-sod1-mutant-motor","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-therapeutics-ad-rna-therapeutics-companies","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-gaps-als-trial-failures","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-cell-types-motor-neurons","link_type":"mentions","strength":0.6,"evidence":"Title match: Motor 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RNA"},{"source_artifact_id":"wiki-entities-c9orf72","link_type":"mentions","strength":0.7,"evidence":"Gene mention: C9ORF72"},{"source_artifact_id":"wiki-cell-types-fus-proteinopathy-neurons","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-ideas-payload-aav-rna-targeting-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-genes-rbfox2","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-proteins-fus","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-proteins-heat-shock-proteins","link_type":"mentions","strength":0.6,"evidence":"Title match: Heat Shock Proteins"},{"source_artifact_id":"wiki-proteins-sod1-protein-expanded","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-experiments-aaav-serotype-lrrk2-gene-therapy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-therapeutics-rna-targeting-therapies-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-therapeutics-aav-gene-therapy-neurodevelopmental-epilepsy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-ideas-payload-fus-protein-targeting-therapy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-diseases","link_type":"mentions","strength":0.6,"evidence":"Title match: Diseases"},{"source_artifact_id":"wiki-proteins-c9orf72","link_type":"mentions","strength":0.7,"evidence":"Gene mention: C9ORF72"},{"source_artifact_id":"wiki-investment-rna-therapeutics","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-ideas-payload-aav-serotype-cns-delivery-optimization-therapy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-mechanisms-als-rna-metabolism-and-proteostasis-failure","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-technologies-crispr-gene-editing","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-mechanisms-als-biomarker-mechanism-map","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-rna-granule-dysfunction-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-mechanisms-als-knowledge-gaps-ranked","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-fus-proteinopathy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-therapeutics-aav-gene-therapy-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-models-aav-lrrk2-gene-delivery-model","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-investment-als-vs-ftd-comparison","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-experiments-aav-serotype-lrrk2-knockdown","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-cell-types-sod1-mutant-motor-neurons","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-cell-types-fus-mutant-motor-neurons","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-mechanisms-rna-metabolism-alzheimers-disease","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-therapeutics-aav-cns-gene-therapy","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-entities-fus","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-mechanisms-als-therapeutic-scorecard","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-technologies-crispr-cas9","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-therapeutics-crispr-gene-editing-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-technologies-aav-vectors","link_type":"mentions","strength":0.7,"evidence":"Gene mention: AAV"},{"source_artifact_id":"wiki-proteins-sod1-protein","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-therapeutics-crispr-gene-editing-parkinsons","link_type":"mentions","strength":0.7,"evidence":"Gene mention: CRISPR"},{"source_artifact_id":"wiki-therapeutics-als-therapeutics","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-als-pathway","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-companies-als-pipeline","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-therapeutics-rnai-therapies-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-therapeutics-sod1-targeting-therapies","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-cell-types-microglia","link_type":"mentions","strength":0.6,"evidence":"Title match: Microglia"},{"source_artifact_id":"wiki-diseases-als-ftd-spectrum","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-cell-types-amyotrophic-lateral-sclerosis-frontotemporal-dementia-neurons","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-therapeutics-als-cure-roadmap","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-als-ftd-unified-pathway","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-rna-metabolism-dysregulation","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-genes-fus","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-entities-sod1-protein","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-entities-fus-protein","link_type":"mentions","strength":0.7,"evidence":"Gene mention: FUS"},{"source_artifact_id":"wiki-therapeutics-rna-targeting-therapy-neurodegeneration","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-diseases-sod1-mutations-als","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-mechanisms-als-molecular-subtypes","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-rna-splicing-4r-tauopathies-comparison","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-mechanisms-als-trial-failure-analysis","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-entities-astrocytes","link_type":"mentions","strength":0.6,"evidence":"Title match: Astrocytes"},{"source_artifact_id":"wiki-institutions-als-clinical-trials-consortium","link_type":"mentions","strength":0.7,"evidence":"Gene mention: ALS"},{"source_artifact_id":"wiki-mechanisms-rna-toxicity","link_type":"mentions","strength":0.7,"evidence":"Gene mention: RNA"},{"source_artifact_id":"wiki-entities-sod1","link_type":"mentions","strength":0.7,"evidence":"Gene mention: SOD1"},{"source_artifact_id":"wiki-cell-types-glial-cells","link_type":"mentions","strength":0.6,"evidence":"Title match: Glial Cells"},{"source_artifact_id":"wiki-cell-types-astrocytes","link_type":"mentions","strength":0.6,"evidence":"Title match: Astrocytes"},{"source_artifact_id":"hypothesis-h-4fabd9ce","link_type":"related","strength":0.85,"evidence":"Hypothesis tested by experiment exp-debate-d1ef3eb9ecab"},{"source_artifact_id":"hypothesis-h-eea667a9","link_type":"related","strength":0.85,"evidence":"Hypothesis tested by experiment exp-debate-d1ef3eb9ecab"},{"source_artifact_id":"hypothesis-h-8196b893","link_type":"related","strength":0.85,"evidence":"Hypothesis tested by experiment exp-debate-d1ef3eb9ecab"},{"source_artifact_id":"hypothesis-h-c463d225","link_type":"related","strength":0.85,"evidence":"Hypothesis tested by experiment exp-debate-d1ef3eb9ecab"},{"source_artifact_id":"hypothesis-h-7e846ceb","link_type":"related","strength":0.85,"evidence":"Hypothesis tested by experiment exp-debate-d1ef3eb9ecab"},{"source_artifact_id":"wiki-genes-hnrnpa2b1","link_type":"mentions","strength":0.7,"evidence":"Gene match: HNRNPA2B1"},{"source_artifact_id":"wiki-genes-setx","link_type":"mentions","strength":0.7,"evidence":"Gene match: SETX"},{"source_artifact_id":"wiki-genes-tardbp","link_type":"mentions","strength":0.7,"evidence":"Gene match: TARDBP"},{"source_artifact_id":"wiki-genes","link_type":"mentions","strength":0.5,"evidence":"Content: TARDBP mentioned 4x in 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wiki/clinical-trials-tofersen-valor"},{"source_artifact_id":"wiki-clinical-trials-genetic-frontotemporal-dementia-nct05653778","link_type":"mentions","strength":0.44999999999999996,"evidence":"Content: TARDBP mentioned 2x in wiki/clinical-trials-genetic-frontotemporal-dementia-nct05653778"},{"source_artifact_id":"wiki-hypotheses-prion-vs-tau-alpha-syn-spread-comparison","link_type":"mentions","strength":0.44999999999999996,"evidence":"Content: TARDBP mentioned 2x in wiki/hypotheses-prion-vs-tau-alpha-syn-spread-comparison"},{"source_artifact_id":"wiki-hypotheses-hyp_575716","link_type":"mentions","strength":0.44999999999999996,"evidence":"Content: TARDBP mentioned 2x in wiki/hypotheses-hyp_575716"},{"source_artifact_id":"wiki-ideas-splice-modulation-therapy-neurodegeneration","link_type":"mentions","strength":0.5,"evidence":"Content: TARDBP mentioned 3x in 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