{"id":"https://openalex.org/W7214959090","doi":"https://doi.org/10.21203/rs.3.rs-11050243/v1","title":"Cas9 nickase-mediated gene editing enables precise large-fragment integration with minimal byproducts","display_name":"Cas9 nickase-mediated gene editing enables precise large-fragment integration with minimal byproducts","publication_year":2026,"publication_date":"2026-09-30","ids":{"openalex":"https://openalex.org/W7214959090","doi":"https://doi.org/10.21203/rs.3.rs-11050243/v1"},"language":"en","primary_location":{"id":"doi:10.21203/rs.3.rs-11050243/v1","doi":"https://doi.org/10.21203/rs.3.rs-11050243/v1","is_oa":true,"landing_page_url":"https://doi.org/10.21203/rs.3.rs-11050243/v1","pdf_url":null,"source":{"id":"https://openalex.org/S4306525896","display_name":"Research Square","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":null,"host_organization_name":null,"host_organization_lineage":[],"host_organization_lineage_names":[],"type":"repository"},"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":"Research Square","raw_type":"posted-content"},"volume":null,"type":"preprint","is_traditional":true,"indexed_in":["crossref"],"open_access":{"is_oa":true,"oa_status":"green","oa_url":"https://doi.org/10.21203/rs.3.rs-11050243/v1","any_repository_has_fulltext":true},"authorships":[{"author_position":"first","author":{"id":"https://openalex.org/A5008419499","display_name":"Sheng Ding","orcid":"https://orcid.org/0000-0002-3354-1263","observed_orcids":["https://orcid.org/0000-0002-3354-1263","https://orcid.org/0000-0002-7525-2144"]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Sheng Ding","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":"https://orcid.org/0000-0002-3354-1263","affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5078192840","display_name":"Kezhang He","orcid":"https://orcid.org/0000-0003-3107-7302","observed_orcids":["https://orcid.org/0000-0003-3107-7302"]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Kezhang He","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":"https://orcid.org/0000-0003-3107-7302","affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5101222163","display_name":"Qiaomei Xue","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Qiaomei Xue","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5153665285","display_name":"Pengqi Wang","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Pengqi Wang","raw_affiliation_strings":["Tsinghua university"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Tsinghua university","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5068570344","display_name":"Qiliang Shao","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Qiliang Shao","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5108883945","display_name":"Weizhe Ding","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Weizhe Ding","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5153656997","display_name":"Yuxia Zhang","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Yuxia Zhang","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5153569503","display_name":"Dan Wang","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Dan Wang","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]},{"author_position":"last","author":{"id":"https://openalex.org/A5102217332","display_name":"Tianhua Ma","orcid":"https://orcid.org/0000-0002-4707-263X","observed_orcids":["https://orcid.org/0000-0002-4707-263X"]},"institutions":[{"id":"https://openalex.org/I99065089","display_name":"Tsinghua University","ror":"https://ror.org/03cve4549","country_code":"CN","type":"education","lineage":["https://openalex.org/I99065089"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Tianhua Ma","raw_affiliation_strings":["Tsinghua University"],"raw_orcid":"https://orcid.org/0000-0002-4707-263X","affiliations":[{"raw_affiliation_string":"Tsinghua University","institution_ids":["https://openalex.org/I99065089"]}]}],"institutions":[],"countries_distinct_count":1,"institutions_distinct_count":1,"corresponding_author_ids":[],"corresponding_institution_ids":["https://openalex.org/I99065089"],"apc_list":null,"apc_paid":null,"fwci":null,"has_fulltext":false,"cited_by_count":0,"citation_normalized_percentile":null,"cited_by_percentile_year":null,"biblio":{"volume":null,"issue":null,"first_page":null,"last_page":null},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T10878","display_name":"CRISPR and Genetic Engineering","score":1.0,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},"topics":[{"id":"https://openalex.org/T10878","display_name":"CRISPR and Genetic Engineering","score":1.0,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T12610","display_name":"RNA regulation and disease","score":0.0,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T10123","display_name":"DNA Repair Mechanisms","score":0.0,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}}],"keywords":[{"id":"https://openalex.org/keywords/cas9-nickase","display_name":"Cas9 nickase","score":0.9629999995231628},{"id":"https://openalex.org/keywords/genome-editing","display_name":"genome editing","score":0.5709999799728394},{"id":"https://openalex.org/keywords/homology-directed-repair","display_name":"homology-directed repair","score":0.7549999952316284},{"id":"https://openalex.org/keywords/prime-editing","display_name":"prime editing","score":0.949999988079071},{"id":"https://openalex.org/keywords/cell-cycle-arrest","display_name":"cell cycle arrest","score":0.4830000102519989}],"concepts":[{"id":"https://openalex.org/C119054055","wikidata":"https://www.wikidata.org/wiki/Q1576681","display_name":"Indel","level":5,"score":0.6658999919891357},{"id":"https://openalex.org/C70721500","wikidata":"https://www.wikidata.org/wiki/Q177005","display_name":"Computational biology","level":1,"score":0.6248000264167786},{"id":"https://openalex.org/C144501496","wikidata":"https://www.wikidata.org/wiki/Q5533489","display_name":"Genome editing","level":4,"score":0.5674999952316284},{"id":"https://openalex.org/C141231307","wikidata":"https://www.wikidata.org/wiki/Q7020","display_name":"Genome","level":3,"score":0.5299000144004822},{"id":"https://openalex.org/C552990157","wikidata":"https://www.wikidata.org/wiki/Q7430","display_name":"DNA","level":2,"score":0.5248000025749207},{"id":"https://openalex.org/C41008148","wikidata":"https://www.wikidata.org/wiki/Q21198","display_name":"Computer science","level":0,"score":0.4918999969959259},{"id":"https://openalex.org/C104317684","wikidata":"https://www.wikidata.org/wiki/Q7187","display_name":"Gene","level":2,"score":0.4823000133037567},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.4821999967098236},{"id":"https://openalex.org/C98108389","wikidata":"https://www.wikidata.org/wiki/Q412563","display_name":"CRISPR","level":3,"score":0.4666000008583069},{"id":"https://openalex.org/C54355233","wikidata":"https://www.wikidata.org/wiki/Q7162","display_name":"Genetics","level":1,"score":0.44190001487731934},{"id":"https://openalex.org/C165525559","wikidata":"https://www.wikidata.org/wiki/Q224180","display_name":"Homology (biology)","level":3,"score":0.3961000144481659},{"id":"https://openalex.org/C197077220","wikidata":"https://www.wikidata.org/wiki/Q720988","display_name":"Human genome","level":4,"score":0.38269999623298645},{"id":"https://openalex.org/C2909909245","wikidata":"https://www.wikidata.org/wiki/Q412550","display_name":"Integrases","level":4,"score":0.3668000102043152},{"id":"https://openalex.org/C2778012447","wikidata":"https://www.wikidata.org/wiki/Q1034415","display_name":"Scope (computer science)","level":2,"score":0.3635999858379364},{"id":"https://openalex.org/C74054111","wikidata":"https://www.wikidata.org/wiki/Q5891445","display_name":"Homology directed repair","level":5,"score":0.35600000619888306},{"id":"https://openalex.org/C51679486","wikidata":"https://www.wikidata.org/wiki/Q380546","display_name":"DNA sequencing","level":3,"score":0.349700003862381},{"id":"https://openalex.org/C2910714999","wikidata":"https://www.wikidata.org/wiki/Q1576681","display_name":"INDEL Mutation","level":5,"score":0.3046000003814697},{"id":"https://openalex.org/C132455925","wikidata":"https://www.wikidata.org/wiki/Q16965677","display_name":"Cas9","level":4,"score":0.2992999851703644},{"id":"https://openalex.org/C44667518","wikidata":"https://www.wikidata.org/wiki/Q12321","display_name":"Palindrome","level":4,"score":0.2865000069141388},{"id":"https://openalex.org/C189206191","wikidata":"https://www.wikidata.org/wiki/Q222046","display_name":"Genomics","level":4,"score":0.28369998931884766},{"id":"https://openalex.org/C134935766","wikidata":"https://www.wikidata.org/wiki/Q210538","display_name":"DNA repair","level":3,"score":0.2565999925136566}],"mesh":[],"locations_count":1,"locations":[{"id":"doi:10.21203/rs.3.rs-11050243/v1","doi":"https://doi.org/10.21203/rs.3.rs-11050243/v1","is_oa":true,"landing_page_url":"https://doi.org/10.21203/rs.3.rs-11050243/v1","pdf_url":null,"source":{"id":"https://openalex.org/S4306525896","display_name":"Research Square","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":null,"host_organization_name":null,"host_organization_lineage":[],"host_organization_lineage_names":[],"type":"repository"},"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":"Research Square","raw_type":"posted-content"}],"best_oa_location":{"id":"doi:10.21203/rs.3.rs-11050243/v1","doi":"https://doi.org/10.21203/rs.3.rs-11050243/v1","is_oa":true,"landing_page_url":"https://doi.org/10.21203/rs.3.rs-11050243/v1","pdf_url":null,"source":{"id":"https://openalex.org/S4306525896","display_name":"Research Square","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":null,"host_organization_name":null,"host_organization_lineage":[],"host_organization_lineage_names":[],"type":"repository"},"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"acceptedVersion","is_accepted":true,"is_published":false,"raw_source_name":"Research Square","raw_type":"posted-content"},"sustainable_development_goals":[],"study_designs":[],"awards":[],"funders":[],"has_content":{"grobid_xml":false,"pdf":false},"content_urls":null,"referenced_works_count":0,"referenced_works":[],"related_works":["https://openalex.org/W2754552929","https://openalex.org/W2777812270","https://openalex.org/W7170058909","https://openalex.org/W4361279075","https://openalex.org/W3003472449","https://openalex.org/W3119556499","https://openalex.org/W3021669023","https://openalex.org/W2991777599","https://openalex.org/W4327676756","https://openalex.org/W3087131783"],"abstract_inverted_index":{"Abstract":[0],"Precise":[1],"installation":[2,85],"of":[3,13,133,193],"large":[4],"DNA":[5,84,134,195],"sequences":[6],"remains":[7],"constrained":[8],"by":[9,111],"the":[10,113,188],"core":[11],"logic":[12],"current":[14],"genome-editing":[15],"platforms.":[16],"Canonical":[17],"homology-directed":[18],"repair":[19,115],"(HDR)":[20],"relies":[21],"on":[22,33],"double-strand":[23,93],"breaks":[24],"and":[25,36,64,82,108,125,143,155,163,186,190],"long":[26,95],"homology":[27,96],"arms,":[28,97],"whereas":[29],"prime-editing-based":[30],"approaches":[31],"depend":[32],"pegRNA-encoded":[34],"information":[35],"reverse":[37,98],"transcription.":[38],"Here":[39],"we":[40],"report":[41],"Nickase-mediated":[42],"Gene":[43],"Editing":[44],"(NKGE),":[45],"a":[46,179],"paired-nick":[47],"strategy":[48],"for":[49,165,182],"precise":[50,131,183],"genome":[51,184],"writing":[52,185],"that":[53,78],"is":[54],"mechanistically":[55],"distinct":[56,180],"from":[57],"both":[58],"paradigms.":[59],"NKGE":[60,129,147,177],"employs":[61],"Cas9(H840A)":[62],"nickases":[63],"paired":[65],"sgRNAs":[66],"to":[67,137,167],"generate":[68],"spatially":[69],"defined":[70],"target-site":[71],"nicks,":[72],"thereby":[73],"creating":[74],"localized":[75],"single-stranded":[76],"regions":[77],"support":[79],"donor":[80],"capture":[81],"seamless":[83],"using":[86],"only":[87],"~17-bp":[88],"terminal":[89],"homology,":[90],"without":[91],"requiring":[92],"breaks,":[94],"transcriptase,":[99],"recombinases,":[100],"or":[101],"transposases.":[102],"This":[103],"design":[104],"actively":[105],"circumvents":[106],"indel":[107],"off-target":[109,145],"formation":[110],"leveraging":[112],"high-fidelity":[114],"pathways":[116],"associated":[117],"with":[118,140],"single-strand":[119],"nicks.":[120],"Across":[121],"multiple":[122],"genomic":[123],"loci":[124],"human":[126],"cell":[127],"lines,":[128],"supported":[130],"insertion":[132],"fragments":[135],"up":[136],"10":[138],"kb":[139],"minimal":[141],"indels":[142],"negligible":[144],"integration.":[146],"also":[148],"retained":[149],"substantial":[150],"activity":[151],"under":[152,170],"cell-cycle":[153],"arrest":[154],"consistently":[156],"outperformed":[157],"HDR,":[158],"prime":[159],"editing,":[160],"Prime":[161],"Assembly,":[162],"QuadPE":[164],"medium-":[166],"large-scale":[168],"insertions":[169],"matched":[171],"conditions.":[172],"Together,":[173],"these":[174],"findings":[175],"establish":[176],"as":[178],"route":[181],"broaden":[187],"mechanistic":[189],"practical":[191],"scope":[192],"therapeutic":[194],"installation.":[196]},"counts_by_year":[],"updated_date":"2026-10-31T12:09:11.096373","created_date":"2026-10-01T00:00:00"}
