title: Highly efficient gene tagging in the bryophyte Physcomitrella patens using the tobacco (Nicotiana tabacum) Tnt1 retrotransposon.
abstract: Because of its highly efficient homologous recombination, the moss Physcomitrella patens is a model organism particularly suited for reverse genetics, but this inherent characteristic limits forward genetic approaches. Here, we show that the tobacco (Nicotiana tabacum) retrotransposon Tnt1 efficiently transposes in P. patens, being the first retrotransposon from a vascular plant reported to transpose in a bryophyte. Tnt1 has a remarkable preference for insertion into genic regions, which makes it particularly suited for gene mutation. In order to stabilize Tnt1 insertions and make it easier to select for insertional mutants, we have developed a two-component system where a mini-Tnt1 with a retrotransposition selectable marker can only transpose when Tnt1 proteins are co-expressed from a separate expression unit. We present a new tool with which to produce insertional mutants in P. patens in a rapid and straightforward manner that complements the existing molecular and genetic toolkit for this model species.
authors.affiliation: Center for Research in Agricultural Genomics, CRAG (CSIC-IRTA-UAB-UB), Campus UAB, Cerdanyola del Vallès, 08193, Barcelona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Vives
authors.affiliation: INRA AgroParisTech, IJPB, UMR 1318, INRA centre de Versailles, route de Saint Cyr, 78026, Versailles Cedex, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Charlot
authors.affiliation: INRA AgroParisTech, IJPB, UMR 1318, INRA centre de Versailles, route de Saint Cyr, 78026, Versailles Cedex, France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Mhiri
authors.affiliation: Center for Research in Agricultural Genomics, CRAG (CSIC-IRTA-UAB-UB), Campus UAB, Cerdanyola del Vallès, 08193, Barcelona, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Contreras
authors.affiliation: INRA AgroParisTech, IJPB, UMR 1318, INRA centre de Versailles, route de Saint Cyr, 78026, Versailles Cedex, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Daniel
authors.affiliation: INRA AgroParisTech, IJPB, UMR 1318, INRA centre de Versailles, route de Saint Cyr, 78026, Versailles Cedex, France.
authors.forename: Aline
authors.initials: A
authors.lastname: Epert
authors.affiliation: Department of Genetics, Cell Biology & Development and Center for Genome Engineering, University of Minnesota, Minneapolis, MN, 55455, USA.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.affiliation: INRA AgroParisTech, IJPB, UMR 1318, INRA centre de Versailles, route de Saint Cyr, 78026, Versailles Cedex, France.
authors.forename: Marie-Angèle
authors.initials: MA
authors.lastname: Grandbastien
authors.affiliation: INRA AgroParisTech, IJPB, UMR 1318, INRA centre de Versailles, route de Saint Cyr, 78026, Versailles Cedex, France. fabien.nogue@versailles.inra.fr.
authors.forename: Fabien
authors.initials: F
authors.lastname: Nogué
authors.affiliation: Center for Research in Agricultural Genomics, CRAG (CSIC-IRTA-UAB-UB), Campus UAB, Cerdanyola del Vallès, 08193, Barcelona, Spain. josep.casacuberta@cragenomica.es.
authors.forename: Josep M
authors.initials: JM
authors.lastname: Casacuberta
mesh_terms: D001483:Base Sequence; D019068:Bryopsida / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D005821:Genetic Techniques; D016254:Mutagenesis, Insertional / Q000235:genetics; D011110:Polymorphism, Genetic; D018626:Retroelements / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics*; D014158:Transcription, Genetic; D014170:Transformation, Genetic
chemical_list: D018626:Retroelements
keywords: 
  Physcomitrella patens
  ; Retrotransposon; Tnt1; gene tagging; mutant population