Rapid and Efficient Suppression of Gene Expression in a Single-Cell Model System, <i>Ceratopteris richardii</i>

Stout, Stephen C.; Clark, Gregory B.; Archer-Evans, Sarah; Roux, Stanley J.

doi:10.1104/pp.016949

Cited by 45 publications

(30 citation statements)

References 22 publications

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“…DNA methylation was detected in the 5' UTR and coding regions of NEO1, but not in the 3' downstream region, suggesting that the DNAi effect was not transmitted to neighboring genes. Stout et al 3 developed an RNAi method in fern using the direct delivery of dsRNA into spores through imbibitions, and it was suggested that a gene silencing system by RNAi machinery exists in Ceratopteris richardii. 3 Indeed, we found genes involved in small RNA biogenesis and epigenetic regulation, such as HDAC, in an Adiantum EST database (http://togodb.dbcls.jp/acest/).…”

Section: Discussionmentioning

confidence: 99%

“…Stout et al 3 developed an RNAi method in fern using the direct delivery of dsRNA into spores through imbibitions, and it was suggested that a gene silencing system by RNAi machinery exists in Ceratopteris richardii. 3 Indeed, we found genes involved in small RNA biogenesis and epigenetic regulation, such as HDAC, in an Adiantum EST database (http://togodb.dbcls.jp/acest/). 16 Ferns may have an RNAi-dependent epigenetic regulation system similar to higher plants, because many of the conserved genes for RNAi systems are present in fern species.…”

Section: Discussionmentioning

confidence: 99%

“…2 In ferns, RNAi in germinating spores provides a simple singlecell system to study gene function. 3 Transcriptional gene silencing and de novo DNA methylation of a transgene-homologous promoter occur if the dsRNA contains promoter sequences. Indeed, an RNAi component is involved in the de novo DNA methylation at cytosine residues in a phenomenon known as RNA-directed DNA methylation.…”

Section: Introductionmentioning

confidence: 99%

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Epigenetic memory of DNAi associated with cytosine methylation and histone modification in fern

Tsuboi

Sutoh

Wada

2012

Plant Signaling & Behavior

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Gene silencing technology, such as rNa interference (rNai), is commonly used to reduce gene expression in plant cells, and exogenous double-stranded rNa (dsrNa) can induce gene silencing in higher plants. Previously, we showed that the delivery of double-stranded DNa (dsDNa) fragments, such as Pcr products of an endogenous gene sequence, into fern (Adiantum capillus-veneris) gametophytic cells induces a sequence-specific gene silencing that we termed DNai. In this study, we used a neochrome 1 gene (NEO1) that mediates both red light-induced chloroplast movement and phototropism as a model of DNai and confirmed that the NEO1 function was suppressed by the repression of the NEO1 gene. Interestingly, the gene silencing effect by DNai was found in the progeny. cytosine methylation was detected in the NEO1-silenced lines. The DNa modifications was present in the transcriptional region of NEO1, but no differences between wild type and the silenced lines were found in the downstream region of NEO1. Our data suggest that the DNai gene silencing effect that was inherited throughout the next generation is regulated by epigenetic modification. Furthermore, the histone deacetylase inhibitor, trichostatin a (TSa), recovered the expression and function of NEO1 in the silenced lines, suggesting that histone deacetylation is essential for the direct suppression of target genes by DNai.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Epigenetic memory of DNAi associated with cytosine methylation and histone modification in fern

Tsuboi

Sutoh

Wada

2012

Plant Signaling & Behavior

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“…Additionally, in plants, RNAi transgenes can be inherited in subsequent generations (Chuang and Meyerowitz, 2000), creating a population of transgenic plants with reduced expression of the gene of interest. In ferns, RNAi in regenerating spores provides a simple single-cell system to study gene function (Klink and Wolniak, 2000;Stout et al, 2003). Here, we show that P. patens also uses RNAi for gene silencing.…”

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confidence: 94%

RNA Interference in the Moss Physcomitrella patens

2003

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The moss Physcomitrella patens performs efficient homologous recombination, which allows for the study of individual gene function by generating gene disruptions. Yet, if the gene of study is essential, gene disruptions cannot be isolated in the predominantly haploid P. patens. Additionally, disruption of a gene does not always generate observable phenotypes due to redundant functions from related genes. However, RNA interference (RNAi) can provide mutants for both of these situations. We show that RNAi disrupts gene expression in P. patens, adding a significant tool for the study of plant gene function. To assay for RNAi in moss, we constructed a line (NLS-4) expressing a nuclearly localized green fluorescent protein (GFP):␤-glucuronidase (GUS) fusion reporter protein. We targeted the reporter protein with two RNAi constructs, GUSRNAi and GFP-RNAi, expressed transiently by particle bombardment. Transformed protonemal cells are marked by cobombardment with dsRed2, which diffuses between the nucleus and cytoplasm. Cells transformed with control constructs have nuclear/cytoplasmic red fluorescence and nuclear green fluorescence. In cells transformed with GUS-RNAi or GFP-RNAi constructs, the nuclear green fluorescence was reduced on average 9-fold as soon as 48 h after transformation. Moreover, isolated lines of NLS-4 stably transformed with GUS-RNAi construct have silenced nuclear GFP, indicating that RNAi is propagated stably. Thus, RNAi adds a powerful tool for functional analysis of plant genes in moss.

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“…The 3,840 cDNA clones corresponding to the EST library described by Stout et al (2003) and deposited in the EST database (GenBank accession nos. BE640669-BE643506 and BQ086920-BQ087668) were amplified by PCR with SP6 and T7 vector-specific primers in 96-well plate format.…”

Section: Microarray Constructionmentioning

confidence: 99%

Profile and Analysis of Gene Expression Changes during Early Development in Germinating Spores of Ceratopteris richardii

et al. 2005

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Analysis of an expressed sequence tag library with more than 5,000 sequences from spores of the fern Ceratopteris richardii reveals that more than 3,900 of them represent distinct genes, and almost 70% of these have significant similarity to Arabidopsis (Arabidopsis thaliana) genes. Eight genes are common between three very different dormant plant systems, Ceratopteris spores, Arabidopsis seeds, and Arabidopsis pollen. We evaluated the pattern of mRNA abundance over the first 48 h of spore development using a microarray of cDNAs representing 3,207 distinct genes of C. richardii and determined the relative levels of RNA abundance for 3,143 of these genes using a Bayesian method of statistical analysis. More than 900 of them (29%) show a significant change between any of the five time points analyzed, and these have been annotated based on their sequence similarity with the Arabidopsis proteome. Novel data arising from these analyses identify genes likely to be critical for the germination and subsequent early development of diverse cells and tissues emerging from dormancy.

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Rapid and Efficient Suppression of Gene Expression in a Single-Cell Model System, Ceratopteris richardii

Cited by 45 publications

References 22 publications

Epigenetic memory of DNAi associated with cytosine methylation and histone modification in fern

Epigenetic memory of DNAi associated with cytosine methylation and histone modification in fern

RNA Interference in the Moss Physcomitrella patens

Profile and Analysis of Gene Expression Changes during Early Development in Germinating Spores of Ceratopteris richardii

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