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dc.description.sponsorshipGefördert durch den Publikationsfonds der Universität Kassel
dc.rightsUrheberrechtlich geschützt
dc.titleA simple retroelement based knock-down system in Dictyosteliumeng
dcterms.abstractCharacteristics of DIRS-1 Mediated Knock-Downs __ We have previously shown that the most abundant Dictyostelium discoideum retroelement DIRS-1 is suppressed by RNAi mechanisms. Here we provide evidence that both inverted terminal repeats have strong promoter activity and that bidirectional expression apparently generates a substrate for Dicer. A cassette containing the inverted terminal repeats and a fragment of a gene of interest was sufficient to activate the RNAi response, resulting in the generation of ~21 nt siRNAs, a reduction of mRNA and protein expression of the respective endogene. Surprisingly, no transitivity was observed on the endogene. This was in contrast to previous observations, where endogenous siRNAs caused spreading on an artificial transgene. Knock-down was successful on seven target genes that we examined. In three cases a phenotypic analysis proved the efficiency of the approach. One of the target genes was apparently essential because no knock-out could be obtained; the RNAi mediated knock-down, however, resulted in a very slow growing culture indicating a still viable reduction of gene expression.eng
dcterms.abstractAdvantages of the DIRS-1–RNAi System __ The knock-down system required a short DNA fragment (~400 bp) of the target gene as an initial trigger. Further siRNAs were generated by RdRPs since we have shown some siRNAs with a 5’-triphosphate group. Extrachromosomal vectors facilitate the procedure and allowed for molecular and phenotypic analysis within one week. The system provides an efficient and rapid method to reduce protein levels including those of essential genes.eng
dcterms.accessRightsopen access
dcterms.alternativeFurther insights into RNA interference mechanismseng
dcterms.bibliographicCitationIn: PLoS one. - Lawrence, Kan. : PLoS, 2015, 10 (6), e0131271, 1-17
dcterms.creatorFriedrich, Michael
dcterms.creatorMeier, Doreen
dcterms.creatorSchuster, Isabelle
dcterms.creatorNellen, Wolfgang
dc.publisher.placeLawrence, Kan.
dcterms.source.journalPLoS one
dcterms.source.pageinfoS. 1-17

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