RNA interference by small hairpin RNAs synthesised under control of the human 7S K RNA promoter.
Small interfering RNAs (siRNAs) represent RNA duplexes of 21 nucleotides in length that inhibit gene expression. We have used the human gene-external 7S K RNA promoter for synthesis of short hairpin RNAs (shRNAs) which efficiently target human lamin mRNA via RNA interference (RNAi). Here we demonstrate that orientation of the target sequence within the shRNA construct is important for interference. Furthermore, effective interference also depends on the length and/or structure of the shRNA. Evidence is presented that the human 7S K promoter is more active in vivo than other gene-external promoters, such as the human U6 small nuclear RNA (snRNA) gene promoter.
Koper-Emde
Dorota
D
Lehrstuhl für Biochemie I, Fakultät für Chemie, Ruhr-Universität Bochum, D-44780 Bochum, Germany.
Herrmann
Lutz
L
Sandrock
Björn
B
Benecke
Bernd-Joachim
BJ
D006367:HeLa Cells; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference / Q000502:physiology*; D020733:RNA, Small Cytoplasmic / Q000096:biosynthesis* / Q000235:genetics; D012342:RNA, Small Nuclear / Q000096:biosynthesis* / Q000235:genetics; D018271:Signal Recognition Particle / Q000096:biosynthesis* / Q000235:genetics
C115990:7SL RNA; D020733:RNA, Small Cytoplasmic; D012342:RNA, Small Nuclear; D018271:Signal Recognition Particle; C405486:U6 small nuclear RNA