Presence of an intron in inverted repeat constructs does not necessarily have an effect on efficiency of post-transcriptional gene silencing

Abstract

The effect of introns on silencing efficiency was tested in inverted repeat constructs of Granule-Bound Starch Synthase (GBSSI) cDNA by comparing the silencing efficiencies induced by inverted repeat constructs with and without introns. No effect could be attributed to the presence of introns indicating that the introns neither enhance nor inhibit post-transcriptional gene silencing. The effect of a spliceable intron in the spacer was studied by comparing constructs harbouring a spliceable or a non-spliceable intron in the spacer. As opposed to the general belief that splicing of an intron increases silencing efficiency, the use of a spliceable intron in the spacer did not result in enhancement of silencing in our experimental system.

Direct Amplification of Intron-Containing Hairpin RNA Construct from Genomic DNA

“…In addition to the ihpRNA constructs, other hpRNA constructs without intron in the spacer were also useful in some systems, showing high gene silencing efficiency (2,3). In these cases, any DNA of appropriate length could be used as a template to amplify hpRNA construct using the DA-ihpRNA method, which might broaden the application of the DA-ihpRNA method to generate constructs for RNAi.…”

Genetic and molecular approaches to valorise protein and fibre in potato

“…Kardal was selected for transformation due to its moderate and relatively stable level of tuber protein content. A. tumerfaciens-mediated transformation was performed as described (Heilersig et al 2006). The developed StNPF1.11 transformants (OE-lines) were coded as OE-x-y, where x denoted the generated series of the transformants and y denoted the unique transformant line number.…”