Loss of sense transgene-induced post-transcriptional gene silencing by sequential introduction of the same transgene sequences in tobacco

Sayaka Hirai, Kouta Takahashi, Tomomi Abiko, Hiroaki Kodama

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

RNA silencing is an epigenetic inhibition of gene expression and is guided by small interfering RNAs. Sense transgene-induced post-transcriptional gene silencing (S-PTGS) occurs in a portion of a transgenic plant population. When a sense transgene encoding a tobacco endoplasmic reticulum ω-3 fatty acid desaturase (NtFAD3) was introduced into tobacco plants, an S-PTGS line, S44, was obtained. Introduction of another copy of the NtFAD3 transgene into S44 plants caused a phenotypic change from S-PTGS to overexpression. Because this change was associated with the methylation of the promoter sequences of the transgene, reduced transcriptional activity may abolish S-PTGS and residual transcription of the sense transgene may account for the overexpression. To clarify whether RNA-directed DNA methylation (RdDM) can repress the transcriptional activity of the S44 transgene locus, we introduced several RdDM constructs targeting the transgene promoter. An RdDM construct harboring a 200-bp-long fragment of promoter sequences efficiently abrogated the generation of NtFAD3 small interfering RNAs in S44 plants. Transcription of the transgene was partially repressed, but the resulting NtFAD3 mRNAs successfully accumulated and an overexpressed phenotype was established. Our results indicate an example in which overexpression of the transgene is established by complex epigenetic interactions among the transgenic loci.

Original languageEnglish
Pages (from-to)1695-1703
Number of pages9
JournalFEBS Journal
Volume277
Issue number7
DOIs
Publication statusPublished - Apr 1 2010

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Tobacco
RNA Interference
Transgenes
Genes
RNA
Transcription
Small Interfering RNA
Fatty Acid Desaturases
Methylation
DNA Methylation
Gene expression
Epigenomics
Messenger RNA
Genetically Modified Plants
Endoplasmic Reticulum

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Loss of sense transgene-induced post-transcriptional gene silencing by sequential introduction of the same transgene sequences in tobacco. / Hirai, Sayaka; Takahashi, Kouta; Abiko, Tomomi; Kodama, Hiroaki.

In: FEBS Journal, Vol. 277, No. 7, 01.04.2010, p. 1695-1703.

Research output: Contribution to journalArticle

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