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Highly efficient gene tagging in the bryophyte Physcomitrella patens using the tobacco ( Nicotiana tabacum ) Tnt1 retrotransposon
Author(s) -
Vives Cristina,
Charlot Florence,
Mhiri Corinne,
Contreras Beatriz,
Daniel Julien,
Epert Aline,
Voytas Daniel F.,
Grandbastien MarieAngèle,
Nogué Fabien,
Casacuberta Josep M.
Publication year - 2016
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/nph.14152
Subject(s) - physcomitrella patens , retrotransposon , biology , nicotiana tabacum , genetics , bryophyte , gene , transposition (logic) , bryopsida , transposable element , insertional mutagenesis , mutant , computational biology , botany , linguistics , philosophy
Summary 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.