{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T07:53:27Z","timestamp":1778658807390,"version":"3.51.4"},"reference-count":44,"publisher":"Proceedings of the National Academy of Sciences","issue":"4","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2011,1,25]]},"abstract":"<jats:p>\n                    DNA transposons have been widely used for transgenesis and insertional mutagenesis in various organisms. Among the transposons active in mammalian cells, the moth-derived transposon\n                    <jats:italic>piggyBac<\/jats:italic>\n                    is most promising with its highly efficient transposition, large cargo capacity, and precise repair of the donor site. Here we report the generation of a hyperactive\n                    <jats:italic>piggyBac<\/jats:italic>\n                    transposase. The active transposition of\n                    <jats:italic>piggyBac<\/jats:italic>\n                    in multiple organisms allowed us to screen a transposase mutant library in yeast for hyperactive mutants and then to test candidates in mouse ES cells. We isolated 18 hyperactive mutants in yeast, among which five were also hyperactive in mammalian cells. By combining all mutations, a total of 7 aa substitutions, into a single reading frame, we generated a unique hyperactive\n                    <jats:italic>piggyBac<\/jats:italic>\n                    transposase with 17-fold and ninefold increases in excision and integration, respectively. We showed its applicability by demonstrating an increased efficiency of generation of transgene-free mouse induced pluripotent stem cells. We also analyzed whether this hyperactive\n                    <jats:italic>piggyBac<\/jats:italic>\n                    transposase affects the genomic integrity of the host cells. The frequency of footprints left by the hyperactive\n                    <jats:italic>piggyBac<\/jats:italic>\n                    transposase was as low as WT transposase (~1%) and we found no evidence that the expression of the transposase affects genomic integrity. This hyperactive\n                    <jats:italic>piggyBac<\/jats:italic>\n                    transposase expands the utility of the\n                    <jats:italic>piggyBac<\/jats:italic>\n                    transposon for applications in mammalian genetics and gene therapy.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1008322108","type":"journal-article","created":{"date-parts":[[2011,1,4]],"date-time":"2011-01-04T20:45:06Z","timestamp":1294173906000},"page":"1531-1536","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":776,"title":["A hyperactive\n                    <i>piggyBac<\/i>\n                    transposase for mammalian applications"],"prefix":"10.1073","volume":"108","author":[{"given":"Kosuke","family":"Yusa","sequence":"first","affiliation":[{"name":"Wellcome Trust Sanger Institute, Cambridge CB10 1SA, United Kingdom; and"}]},{"given":"Liqin","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, The Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205-2185"}]},{"given":"Meng Amy","family":"Li","sequence":"additional","affiliation":[{"name":"Wellcome Trust Sanger Institute, Cambridge CB10 1SA, United Kingdom; and"}]},{"given":"Allan","family":"Bradley","sequence":"additional","affiliation":[{"name":"Wellcome Trust Sanger Institute, Cambridge CB10 1SA, United Kingdom; and"}]},{"given":"Nancy L.","family":"Craig","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, The Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205-2185"}]}],"member":"341","published-online":{"date-parts":[[2011,1,4]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.3.9.1288"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.24.10824"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(85)90230-2"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.83.8.2579"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/0955-0674(95)80097-2"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80436-5"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.23.24.9189-9207.2003"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth795"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth1002"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1461"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/165.1.243"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature03681"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature03691"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1526"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1163040"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2010.2"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2005.07.013"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0606979103"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0801017105"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkm446"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1323"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/ng.343"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.20508"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2008.41"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(00)80397-4"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/22.25.5767"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrc2523"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/258121a0"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/0027-5107(94)90305-0"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0965-1748(02)00084-X"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2335980100"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0504679102"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0507683103"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg341"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.24.9.4004-4018.2004"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.01500-06"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0798-657"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-9525(00)02104-1"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/153.3.1403"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.103.026229"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.104.035576"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.0020156"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0805638105"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2009.07.011"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.1008322108","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T19:42:15Z","timestamp":1649792535000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.1008322108"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,1,4]]},"references-count":44,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2011,1,25]]}},"alternative-id":["10.1073\/pnas.1008322108"],"URL":"https:\/\/doi.org\/10.1073\/pnas.1008322108","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.8560957.9046056","asserted-by":"object"}]},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,1,4]]},"assertion":[{"value":"2011-01-04","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}