Highly efficient gene tagging in the bryophyte Physcomitrella patens using the tobacco (Nicotiana tabacum) Tnt1 retrotransposon

Cristina Vives, Florence Charlot, Corinne Mhiri, Beatriz Contreras, Julien Daniel, Aline Epert, Daniel F. Voytas, Marie Angèle Grandbastien, Fabien Nogué, Josep M. Casacuberta

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)759-769
Number of pages11
JournalNew Phytologist
Volume212
Issue number3
DOIs
StatePublished - Nov 1 2016

Keywords

  • Physcomitrella patens
  • Retrotransposon
  • Tnt1
  • gene tagging
  • mutant population

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