{"doi":"10.1126/science.adj8543","title":"Mechanism of target site selection by type V-K CRISPR-associated transposases","abstract":"CRISPR-associated transposases (CASTs) repurpose nuclease-deficient CRISPR effectors to catalyze RNA-guided transposition of large genetic payloads. Type V-K CASTs offer potential technology advantages but lack accuracy, and the molecular basis for this drawback has remained elusive. Here, we reveal that type V-K CASTs maintain an RNA-independent, \"untargeted\" transposition pathway alongside RNA-dependent integration, driven by the local availability of TnsC filaments. Using cryo-electron microscopy, single-molecule experiments, and high-throughput sequencing, we found that a minimal, CRISPR-less transpososome preferentially directs untargeted integration at AT-rich sites, with additional local specificity imparted by TnsB. By exploiting this knowledge, we suppressed untargeted transposition and increased type V-K CAST specificity up to 98.1% in cells without compromising on-target integration efficiency. These findings will inform further engineering of CAST systems for accurate, kilobase-scale genome engineering applications.","journal":"Science","year":2023,"id":325221,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":32,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9556,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":552747,"name":"Christopher Acree","orcid":"0000-0002-3958-1055","position":1,"is_corresponding":false},{"id":646122,"name":"Jung-Un Park","orcid":"0000-0002-0269-7142","position":2,"is_corresponding":false},{"id":247317,"name":"Muwen Kong","orcid":"0000-0002-7845-1165","position":3,"is_corresponding":false},{"id":226296,"name":"Tanner Wiegand","orcid":"0000-0002-0528-268X","position":4,"is_corresponding":false},{"id":1042741,"name":"Yanis Luca Pignot","orcid":"0000-0003-1087-285X","position":5,"is_corresponding":false},{"id":351499,"name":"Elizabeth H. Kellogg","orcid":"0000-0002-9664-3179","position":6,"is_corresponding":false},{"id":247324,"name":"Eric C. Greene","orcid":"0000-0002-7387-824X","position":7,"is_corresponding":false},{"id":561678,"name":"Samuel H. Sternberg","orcid":"0000-0001-8240-9114","position":8,"is_corresponding":false},{"id":856108,"name":"Jerrin Thomas George","orcid":"0000-0001-9696-0564","position":0,"is_corresponding":true}],"reference_count":63,"raw_metadata":null,"created_at":"2026-07-19T01:08:22.919783Z","pmid":"37972161","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":[]}