Impact of differential <scp>DNA</scp> methylation on transgene expression in cotton <i>(Gossypium hirsutum L.)</i> events generated by targeted sequence insertion

Verkest, Aurine; Bourout, Stephane; Debaveye, Jurgen; Reynaert, Kristine; Saey, Bernadette; Brande, Ilse van den; D’Halluin, Kathleen

doi:10.1111/pbi.13049

Cited by 5 publications

(3 citation statements)

References 55 publications

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“…High DNA methylation inhibits gene expression, ultimately resulting in gene silencing (Pierard et al, 2010 ; Lou et al, 2014 ). In transgenic cotton, Verkest et al ( 2019 ) find that transgenic plants stably expressing exogenous genes show little or no methylation, whereas plants with silenced exogenous genes show hypermethylation in the promoter and coding regions.…”

Section: Introductionmentioning

confidence: 99%

DNA Methylation Silences Exogenous Gene Expression in Transgenic Birch Progeny

Chen

Yin

et al. 2020

Front. Plant Sci.

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The genetic stability of exogenous genes in the progeny of transgenic trees is extremely important in forest breeding; however, it remains largely unclear. We selected transgenic birch (Betula platyphylla) and its hybrid F1 progeny to investigate the expression stability and silencing mechanism of exogenous genes. We found that the exogenous genes of transgenic birch could be transmitted to their offspring through sexual reproduction. The exogenous genes were segregated during genetic transmission. The hybrid progeny of transgenic birch WT1×TP22 (184) and WT1×TP23 (212) showed higher Bgt expression and greater insect resistance than their parents. However, the hybrid progeny of transgenic birch TP23×TP49 (196) showed much lower Bgt expression, which was only 13.5% of the expression in its parents. To elucidate the mechanism underlying the variation in gene expression between the parents and progeny, we analyzed the methylation rates of Bgt in its promoter and coding regions. The hybrid progeny with normally expressed exogenous genes showed much lower methylation rates (0–29%) than the hybrid progeny with silenced exogenous genes (32.35–45.95%). These results suggest that transgene silencing in the progeny is mainly due to DNA methylation at cytosine residues. We further demonstrated that methylation in the promoter region, rather than in the coding region, leads to gene silencing. We also investigated the relative expression levels of three methyltransferase genes: BpCMT, BpDRM, and BpMET. The transgenic birch line 196 with a silenced Gus gene showed, respectively, 2.54, 9.92, and 4.54 times higher expression levels of BpCMT, BpDRM, and BpMET than its parents. These trends are consistent with and corroborate the high methylation levels of exogenous genes in the transgenic birch line 196. Therefore, our study suggests that DNA methylation in the promoter region leads to silencing of exogenous genes in transgenic progeny of birch.

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Section: Introductionmentioning

confidence: 99%

DNA Methylation Silences Exogenous Gene Expression in Transgenic Birch Progeny

Chen

Yin

et al. 2020

Front. Plant Sci.

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“…These DNA secondary structures are associated with the recruitment of DNA methylation machinery (including de novo methyltransferase) and initiation of DNA methylation (Bestor, 1987; Bestor & Tycko, 1996; O'Hagan et al, 2008; Wei et al, 2012; Gentry & Meyer, 2013; Gentry & Hennig, 2016). Thus, it is conceivable that a common DNA secondary structure mediated by the recombinases rather than specific sequence elements could be responsible for triggering and spreading the DNA methylation (Bestor & Tycko, 1996; Gentry & Meyer, 2013; Verkest et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

DNA methylation occurring in Cre‐expressing cells inhibits loxP recombination and silences loxP‐sandwiched genes

et al. 2021

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The low DNA recombination efficiency of site-specific recombinase systems in plants limits their application; however, the underlying mechanism is unknown.We evaluate the gene deletion performance of four recombinase systems (Cre/loxP, Flp/ FRT, KD/KDRT and B3/B3RT) in tobacco where the recombinases are under the control of germline-specific promoters.We find that the expression of these recombinases results mostly in gene silencing rather than gene deletion. Using the Cre/loxP system as a model, we reveal that the region flanked by loxP sites (floxed) is hypermethylated, which prevents floxed genes from deletion while silencing the expression of the genes. We further show CG methylation alone in the recombinase binding element of the loxP site is unable to impede gene deletion; instead, CHH methylation in the crossover region is required to inhibit loxP recombination.Our study illustrates the important role of recombinase-induced DNA methylation in the inhibition of site-specific DNA recombination and uncovers the mechanism underlying recombinase-associated gene silence in plants.

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“…High DNA methylation inhibits gene expression, ultimately resulting in gene silencing (Pierard et al, 2010;Lou et al, 2014). In transgenic cotton, Verkest et al (2019) find that transgenic plants stably expressing exogenous genes show little or no methylation, whereas plants with silenced exogenous genes show hypermethylation in the promoter and coding regions.…”

Section: Condensation Of Chromatin or Degradation Of Transcripts Bymentioning

confidence: 99%

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Impact of differential DNA methylation on transgene expression in cotton (Gossypium hirsutum L.) events generated by targeted sequence insertion

Cited by 5 publications

References 55 publications

DNA Methylation Silences Exogenous Gene Expression in Transgenic Birch Progeny

DNA Methylation Silences Exogenous Gene Expression in Transgenic Birch Progeny

DNA methylation occurring in Cre‐expressing cells inhibits loxP recombination and silences loxP‐sandwiched genes

Data_Sheet_1.docx

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