{"doi":"10.1016/j.ymthe.2022.01.005","title":"A flexible split prime editor using truncated reverse transcriptase improves dual-AAV delivery in mouse liver","abstract":"Prime editor (PE) has tremendous promise for gene therapy. However, it remains a challenge to deliver PE (>6.3 kb) in vivo. Although PE can be split into two fragments and delivered using dual adeno-associated viruses (AAVs), choice of split sites within Cas9-which affects editing efficiency-is limited due to the large size of PE. Furthermore, overexpressing reverse transcriptase in mammalian cells might disrupt translation termination via its RNase H domain. Here, we developed a compact PE without the RNase H domain that showed editing comparable with full-length PE. With compact PE, we used a Cas9 split site (Glu 573) that supported robust editing in cells (up to 93% of full-length PE) and in mouse liver. We then demonstrated that split-cPE573 delivered by dual-AAV8 efficiently mediated a 3-bp TGA insertion in the Pcsk9 gene in mouse liver. Compact PE without the RNase H domain abolished its binding to peptidyl release factor 1 (eRF1) and mitigated the stop codon readthrough effect observed with full-length PE. This study identifies a compact PE with a flexible split design to advance utility of prime editing in vivo.","journal":"Molecular Therapy","year":2022,"id":233924,"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":111,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9543,"is_data_producer":false,"deposit_databanks":null,"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":362071,"name":"Shun‐Qing Liang","orcid":"0000-0003-4664-6029","position":1,"is_corresponding":false},{"id":843914,"name":"Bin Liu","orcid":"0000-0002-3168-8456","position":2,"is_corresponding":false},{"id":306037,"name":"Pengpeng Liu","orcid":"0000-0002-4891-8780","position":3,"is_corresponding":false},{"id":849041,"name":"Suet-Yan Kwan","orcid":null,"position":4,"is_corresponding":false},{"id":588037,"name":"Scot A. Wolfe","orcid":"0000-0002-7042-201X","position":5,"is_corresponding":false},{"id":265835,"name":"Wen Xue","orcid":"0000-0002-9797-8042","position":6,"is_corresponding":false},{"id":620176,"name":"Chunwei Zheng","orcid":"0000-0002-2084-5567","position":0,"is_corresponding":true}],"reference_count":30,"raw_metadata":null,"created_at":"2026-07-19T00:21:31.237180Z","pmid":"34998953","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":[]}