Journal article
The key role of terminators on the expression and post‐transcriptional gene silencing of transgenes
The Plant Journal, Vol.104(1), pp.96-112
2020
PMID: 32603508
Abstract
Transgenes 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.
Details
- Title
- The key role of terminators on the expression and post‐transcriptional gene silencing of transgenes
- Authors
- Felipe de Felippes (Corresponding Author) - Queensland University of Technology (QUT)M McHale (Author) - National University of Ireland Galway (NUIG)Rachel L. Doran (Author) - Queensland University of Technology (QUT)Sally Roden (Author) - Queensland University of Technology (QUT)Andrew Eamens (Author) - University of Newcastle AustraliaE. Jean Finnegan (Author) - CSIRO Agriculture and FoodPeter M. Waterhouse (Corresponding Author) - Queensland University of Technology (QUT)
- Publication details
- The Plant Journal, Vol.104(1), pp.96-112
- Publisher
- Wiley-Blackwell Publishing Ltd.
- DOI
- 10.1111/tpj.14907
- ISSN
- 1365-313X
- PMID
- 32603508
- Grant note
- Australian Research Council (FL160100155)
- Organisation Unit
- School of Health and Behavioural Sciences - Legacy; University of the Sunshine Coast, Queensland; School of Health - Biomedicine
- Language
- English
- Record Identifier
- 99656593002621
- Output Type
- Journal article
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- Domestic collaboration
- International collaboration
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- Plant Sciences
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Source: InCites