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Updated: May 3, 2026

Screening and Identification of RNA Silencing Suppressors from Secreted Effectors of Plant Pathogens
Published on: February 3, 2020
Detection and investigation of transitive gene silencing in plants
Leen Vermeersch1, Nancy De Winne, Ann Depicker
1Department of Plant Systems Biology, VIB, Ghent University, Ghent, Belgium.
Abstract:
RNA-induced post-transcriptional silencing is a common tool in functional gene analysis and its application in crop improvement is widely investigated. However, its specificity might be impaired by off-target silencing as a result of transitivity. Generally transitivity is investigated by the detection of secondary siRNAs; however, these tests fail to demonstrate the siRNA's bioactivity. Here, we describe protocols to detect the occurrence of transitive silencing across an endogene by using a reporter GUS gene. In addition, we provide a setup to test the influence of a sequence of interest present in the primary target on the progression of transitivity.
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