System for stable β-estradiol-inducible gene expression in the moss Physcomitrella patens

PLoS One. 2013 Sep 27;8(9):e77356. doi: 10.1371/journal.pone.0077356. eCollection 2013.

Abstract

Inducible transgene expression provides a useful tool to analyze gene function. The moss Physcomitrellapatens is a model basal land plant with well-developed research tools, including a high efficiency of gene targeting and substantial genomics resources. However, current systems for controlled transgene expression remain limited. Here we report the development of an estrogen receptor mediated inducible gene expression system, based on the system used in flowering plants. After identifying the appropriate promoters to drive the chimeric transducer, we succeeded in inducing transcription over 1,000-fold after 24 h incubation with β-estradiol. The P. patens system was also effective for high-level long-term induction of gene expression; transcript levels of the activated gene were maintained for at least seven days on medium containing β-estradiol. We also established two potentially neutral targeting sites and a set of vectors for reproducible expression of two transgenes. This β-estradiol-dependent system will be useful to test genes individually or in combination, allowing stable, inducible transgenic expression in P. patens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bryopsida / drug effects*
  • Bryopsida / genetics*
  • Dose-Response Relationship, Drug
  • Estradiol / pharmacology*
  • Gene Expression / drug effects
  • Genetic Engineering / methods*
  • Genetic Loci / drug effects
  • Genetic Loci / genetics
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / genetics
  • Recombinant Fusion Proteins / genetics
  • Time Factors
  • Transcriptional Activation / drug effects*
  • Transgenes / genetics

Substances

  • Recombinant Fusion Proteins
  • Estradiol

Grants and funding

This work was supported by the Japan Science and Technology Agency under the Exploratory Research for Advanced Technology program to MH, the Strategic Young Researcher Overseas Visits Program for Accelerating Brain Circulation to RR and MH, and the Ministry of Education, Culture, Sports, Science and Technology to TN and MH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.