
SummaryTransgenes have become essential to modern biology, being an important tool in functional genomic studies and also in the development of biotechnological products. One of the major challenges in the generation of transgenic lines concerns the expression of transgenes, which, compared to endogenes, are particularly susceptible to silencing mediated by small RNAs (sRNAs). Several reasons have been put forward to explain why transgenes often trigger the production of sRNAs, such as the high level of expression induced by commonly used strong constitutive promoters, the lack of introns, and features resembling viral and other exogenous sequences. However, the relative contributions of the different genomic elements with respect to protecting genes from the silencing machinery and their molecular mechanisms remain unclear. Here, we present the results of a mutagenesis screen conceived to identify features involved in the protection of endogenes against becoming a template for the production of sRNAs. Interestingly, all of the recovered mutants had alterations in genes with proposed function in transcription termination, suggesting a central role of terminators in this process. Indeed, using a GFP reporter system, we show that, among different genetic elements tested, the terminator sequence had the greatest effect on transgene‐derived sRNA accumulation and that a well‐defined poly(A) site might be especially important. Finally, we describe an unexpected mechanism, where transgenes containing certain intron/terminator combinations lead to an increase in the production of sRNAs, which appears to interfere with splicing.
Nicotiana, 570, Transcription, Genetic, Arabidopsis, Plant Science, PTGS, 576, HSP terminator, 1307 Cell Biology, 1311 Genetics, 1110 Plant Science, Genetics, Transgenes, RNA, Small Interfering, transitivity, transcription termination, Terminator Regions, Genetic, Cell Biology, siRNAs, Mutagenesis, silencing, RNA Interference
Nicotiana, 570, Transcription, Genetic, Arabidopsis, Plant Science, PTGS, 576, HSP terminator, 1307 Cell Biology, 1311 Genetics, 1110 Plant Science, Genetics, Transgenes, RNA, Small Interfering, transitivity, transcription termination, Terminator Regions, Genetic, Cell Biology, siRNAs, Mutagenesis, silencing, RNA Interference
| selected citations These citations are derived from selected sources. This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically). | 68 | |
| popularity This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network. | Top 1% | |
| influence This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically). | Top 10% | |
| impulse This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network. | Top 1% |
