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1000 Titel
  • A single amino acid switch converts the Sleeping Beauty transposase into an efficient unidirectional excisionase with utility in stem cell reprogramming
1000 Autor/in
  1. Kesselring, Lisa |
  2. Miskey, Csaba |
  3. Zuliani, Cecilia Ines |
  4. Querques, Irma |
  5. Kapitonov, Vladimir |
  6. Laukó, Andrea |
  7. Fehér, Anita |
  8. Palazzo, Antonio |
  9. Diem, Tanja |
  10. Lustig, Janna |
  11. Sebe, Attila |
  12. Wang, Yongming |
  13. Dinnyés, András |
  14. Izsvak, Zsuzsanna |
  15. Barabas, Orsolya |
  16. Ivics, Zoltan |
1000 Publikationstyp
  1. Artikel |
1000 Online veröffentlicht
  • 2019-11-28
1000 Erschienen in
1000 Quellenangabe
  • 48(1):316-331
1000 FRL-Sammlung
1000 Copyrightjahr
  • 2019
1000 Lizenz
1000 Verlagsversion
  • https://doi.org/10.1093/nar/gkz1119 |
  • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6943129/ |
1000 Ergänzendes Material
  • https://academic.oup.com/nar/article/48/1/316/5644994#supplementary-data |
1000 Publikationsstatus
1000 Begutachtungsstatus
1000 Sprache der Publikation
1000 Abstract/Summary
  • The Sleeping Beauty (SB) transposon is an advanced tool for genetic engineering and a useful model to investigate cut-and-paste DNA transposition in vertebrate cells. Here, we identify novel SB transposase mutants that display efficient and canonical excision but practically unmeasurable genomic re-integration. Based on phylogenetic analyses, we establish compensating amino acid replacements that fully rescue the integration defect of these mutants, suggesting epistasis between these amino acid residues. We further show that the transposons excised by the exc+/int− transposase mutants form extrachromosomal circles that cannot undergo a further round of transposition, thereby representing dead-end products of the excision reaction. Finally, we demonstrate the utility of the exc+/int− transposase in cassette removal for the generation of reprogramming factor-free induced pluripotent stem cells. Lack of genomic integration and formation of transposon circles following excision is reminiscent of signal sequence removal during V(D)J recombination, and implies that cut-and-paste DNA transposition can be converted to a unidirectional process by a single amino acid change.
1000 Sacherschließung
gnd 4071722-7 Gentechnologie
gnd 4233582-6 Transposon
1000 Fächerklassifikation (DDC)
1000 Liste der Beteiligten
  1. https://frl.publisso.de/adhoc/uri/S2Vzc2VscmluZywgTGlzYSAgICA=|https://orcid.org/0000-0002-7566-9556|https://orcid.org/0000-0001-6026-7207|https://orcid.org/0000-0003-1113-2773|https://frl.publisso.de/adhoc/uri/S2FwaXRvbm92LCBWbGFkaW1pciA=|https://frl.publisso.de/adhoc/uri/ICBMYXVrw7MsIEFuZHJlYSAg|https://frl.publisso.de/adhoc/uri/ICAgRmVow6lyLCBBbml0YSAgIA==|https://frl.publisso.de/adhoc/uri/ICBQYWxhenpvLCBBbnRvbmlvICA=|https://frl.publisso.de/adhoc/uri/ICAgRGllbSwgVGFuamEgICAgIA==|https://frl.publisso.de/adhoc/uri/ICBMdXN0aWcsIEphbm5hICAgICAg|https://orcid.org/0000-0001-7814-5144|https://orcid.org/0000-0001-8269-5296|https://frl.publisso.de/adhoc/uri/IERpbm55w6lzLCBBbmRyw6FzICAgICA=|https://orcid.org/0000-0002-2053-2384|https://orcid.org/0000-0002-2873-5872|https://orcid.org/0000-0002-7803-6658
1000 Label
1000 Förderer
  1. Seventh Framework Programme |
1000 Fördernummer
  1. 230675
1000 Förderprogramm
  1. InduStem
1000 Dateien
1000 Förderung
  1. 1000 joinedFunding-child
    1000 Förderer Seventh Framework Programme |
    1000 Förderprogramm InduStem
    1000 Fördernummer 230675
1000 Objektart article
1000 Beschrieben durch
1000 @id frl:6432662.rdf
1000 Erstellt am 2022-03-29T14:27:58.421+0200
1000 Erstellt von 323
1000 beschreibt frl:6432662
1000 Bearbeitet von 317
1000 Zuletzt bearbeitet Wed May 04 09:43:38 CEST 2022
1000 Objekt bearb. Wed May 04 09:43:26 CEST 2022
1000 Vgl. frl:6432662
1000 Oai Id
  1. oai:frl.publisso.de:frl:6432662 |
1000 Sichtbarkeit Metadaten public
1000 Sichtbarkeit Daten public
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