{"doi":"10.1016/j.omtm.2022.08.005","title":"PAM-altering SNP-based allele-specific CRISPR-Cas9 therapeutic strategies for Huntington’s disease","abstract":"Huntington's disease (HD) is caused by an expanded CAG repeat in huntingtin ( HTT ). Since HD is dominant and loss of HTT leads to neurological abnormalities, safe therapeutic strategies require selective inactivation of mutant HTT . Previously, we proposed a concept of CRISPR-Cas9 using mutant-specific PAM sites generated by SNPs to selectively inactivate mutant HTT . Aiming at revealing suitable targets for clinical development, we analyzed the largest HD genotype dataset to identify target PAM-altering SNPs (PAS) and subsequently evaluated their allele specificities. The gRNAs based on the PAM sites generated by rs2857935, rs16843804, and rs16843836 showed high levels of allele specificity in patient-derived cells. Simultaneous use of two gRNAs based on rs2857935-rs16843804 or rs2857935-rs16843836 produced selective genomic deletions in mutant HTT and prevented the transcription of mutant HTT mRNA without impacting the expression of normal counterpart or re-integration of the excised fragment elsewhere in the genome. RNA-seq and off-target analysis confirmed high levels of allele specificity and the lack of recurrent off-targeting. Approximately 60% of HD subjects are eligible for mutant-specific CRISPR-Cas9 strategies of targeting one of these three PAS in conjunction with one non-allele-specific site, supporting high applicability of PAS-based allele-specific CRISPR approaches in the HD patient population.","journal":"Molecular Therapy — Methods & Clinical Development","year":2022,"id":258938,"datarank":0.6511442048777213,"base_score":2.8903717578961645,"endowment":2.8903717578961645,"self_citation_contribution":0.4335557636844247,"citation_network_contribution":0.2175884411932966,"self_endowment_contribution":0.4335557636844247,"citer_contribution":0.2175884411932966,"corpus_percentile":null,"corpus_rank":null,"citation_count":17,"citer_count":15,"citers_with_citation_signal":11,"citers_with_endowment":11,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9453,"is_data_producer":true,"deposit_databanks":{"Dryad":["10.5061/dryad.1ns1rn8vv"]},"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":13565,"name":"Eun Pyo Hong","orcid":"0000-0001-7789-686X","position":1,"is_corresponding":false},{"id":875739,"name":"Seri S. Park","orcid":null,"position":2,"is_corresponding":false},{"id":875166,"name":"Doo Eun Choi","orcid":"0009-0003-4416-2386","position":3,"is_corresponding":false},{"id":911693,"name":"Sophia Zeng","orcid":null,"position":4,"is_corresponding":false},{"id":875168,"name":"Richard Z. Chen","orcid":"0000-0002-3548-5760","position":5,"is_corresponding":false},{"id":142182,"name":"Jong-Min Lee","orcid":"0000-0001-5799-0787","position":6,"is_corresponding":false},{"id":303419,"name":"Jun Wan Shin","orcid":null,"position":0,"is_corresponding":true}],"reference_count":99,"raw_metadata":null,"created_at":"2026-07-19T00:25:47.305414Z","pmid":"36092363","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}