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

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 tran...

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Bibliographic Details
Published in:PloS one 2013-09, Vol.8 (9), p.e77356
Main Authors: Kubo, Minoru, Imai, Akihiro, Nishiyama, Tomoaki, Ishikawa, Masaki, Sato, Yoshikatsu, Kurata, Tetsuya, Hiwatashi, Yuji, Reski, Ralf, Hasebe, Mitsuyasu
Format: Article
Language:English
Subjects:
17β-Estradiol
Biology
Bryopsida - drug effects
Bryopsida - genetics
Deoxyribonucleic acid
DNA
Dose-Response Relationship, Drug
Estradiol - pharmacology
Estrogens
Expression vectors
Flowering
Gene expression
Gene Expression - drug effects
Gene targeting
Genetic Engineering - methods
Genetic Loci - drug effects
Genetic Loci - genetics
Genomes
Genomics
Incubation
Life sciences
Mosses
Mutagenesis
Physcomitrella patens
Promoter Regions, Genetic - drug effects
Promoter Regions, Genetic - genetics
Proteins
Recombinant Fusion Proteins - genetics
RNA polymerase
Sex hormones
Time Factors
Transcription
Transcriptional Activation - drug effects
Transgenes
Transgenes - genetics
Transgenic
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0077356