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However, production requires methods to trans\u2010complement viral proteins, usually through Helper Viruses (HV). The Cre\/loxP system is frequently employed to remove the packaging signal in HV genomes, in order to avoid their encapsidation. However, chronic exposure to the Cre recombinase in packaging cells is detrimental. We have applied the dimerizable Cre system to overcome this limitation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods and Results<\/jats:title><jats:p>Cre was split in two fragments devoid of recombinase function (N\u2010terminal 244 and C\u2010terminal 99 amino\u2010acids). In one version of the system, interaction with both moieties was favored by rapamycin\u2010dependent heterodimerization domains (DiCre). Other version contained only Cre sequences (oCre). We generated packaging cells and HVs expressing the complementary fragments and studied their performance for HC\u2010AdV production. We found that both conformations avoided interference with the growth of packaging cells, and the oCre system was particularly suitable for HC\u2010AdV amplification.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The split\u2010Cre system improves the performance of packaging cells and can reduce the time and cost of HC\u2010AdV amplification up to 30% and 15%, respectively. This may contribute to the standardization of HC\u2010AdV production.<\/jats:p><\/jats:sec>","DOI":"10.1002\/biot.202200227","type":"journal-article","created":{"date-parts":[[2022,12,7]],"date-time":"2022-12-07T16:52:11Z","timestamp":1670431931000},"source":"Crossref","is-referenced-by-count":5,"title":["Application of a split\u2010Cre system for high\u2010capacity adenoviral vector amplification"],"prefix":"10.1002","volume":"18","author":[{"given":"Manuela","family":"Gonzalez\u2010Aparicio","sequence":"first","affiliation":[{"name":"University of Navarra, CIMA, Gene Therapy and Regulation of Gene Expression Program Navarra Health Research Institute (IdiSNA)  Pamplona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Bunuales","sequence":"additional","affiliation":[{"name":"University of Navarra, CIMA, Gene Therapy and Regulation of Gene Expression Program Navarra Health Research Institute (IdiSNA)  Pamplona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"I\u00f1aki Ortiz","family":"de Landazuri","sequence":"additional","affiliation":[{"name":"University of Navarra, CIMA, Gene Therapy and Regulation of Gene Expression Program Navarra Health Research Institute (IdiSNA)  Pamplona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesus","family":"Prieto","sequence":"additional","affiliation":[{"name":"University of Navarra, CIMA, Gene Therapy and Regulation of Gene Expression Program Navarra Health Research Institute (IdiSNA)  Pamplona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6682-2598","authenticated-orcid":false,"given":"Ruben","family":"Hernandez\u2010Alcoceba","sequence":"additional","affiliation":[{"name":"University of Navarra, CIMA, Gene Therapy and Regulation of Gene Expression Program Navarra Health Research Institute (IdiSNA)  Pamplona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,12,12]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/1873\u20103468.13691"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1099\/0022\u20101317\u201036\u20101\u201059"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-021-00356-x"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/1074\u20107613(94)90074\u20104"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21103643"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2013.071"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.24.13565"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.3791\/52894"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2021.01.002"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/89349"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1006\/mthe.2001.0323"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2021.12.001"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.9.4181-4189.2002"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.161269798"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348\u20100421.2005.tb03753.x"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2021.109105"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.240471297"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2013.05.019"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/gt.2011.58"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gng131"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/pbi.12726"},{"issue":"5","key":"e_1_2_10_23_1","first-page":"441","article-title":"Targeted insertion of exogenous DNA into the eukaryotic genome by the Cre recombinase","volume":"2","author":"Sauer B.","year":"1990","journal-title":"The New Biologist"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.5.2509"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02369439"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2002.1406"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkq384"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167\u20104781(89)90016\u2010X"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/27.24.4703"},{"issue":"58","key":"e_1_2_10_30_1","first-page":"87","article-title":"a s tructural v iew of cre\u2010loxp s ite \u2010s pecific r ecombination","volume":"1","author":"Duyne G. 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