Silencing of antibody genes in plants with single-copy transgene inserts as a result of gene dosage effects

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Abstract
The stability of Fab antibody fragment expression during plant development was studied using two homozygous Arabidopsis thaliana lines that contain single copies of the transgenes. These lines exhibited expression characteristics that are typical for homology-based post-transcriptional gene silencing. Their developmental silencing profiles differed markedly, presumably due to the influence of the genomic context on the T-DNAs. In both lines, a clear gene dosage effect could be observed: in contrast to the homozygous lines, derived hemizygous plants accumulated high levels of Fab fragments throughout development. Interestingly, silencing also occurred in double-hemizygous plants, which resulted from a cross between the two homozygous lines and had two copies of each T-DNA at non-allelic positions in their genome. In all cases, down-regulation of the Fab levels was strictly correlated with methylation of cytosine residues in the transcribed regions of the transgenes. Remarkably, this methylation was also found in regions in which the transgenes were non-homologous regions. Finally, the time point of down-regulation depended on the culture conditions and differed for leaves and roots of the same transgenic plant.

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