Light-dependent RNA interference with nucleobase-caged siRNAs.
Within the past years RNA interference (RNAi) has become one of the most valuable tools for post-transcriptional gene silencing. Making RNAi temporally and/or spatially controllable would even enlarge its scope of application. Attaching a light-removable protection group to siRNAs is a very promising approach to achieve this control over RNAi. It has been reported that modifying siRNA nucleobases surrounding the mRNA cleavage site between the 10th and 11th nucleotides successfully suppresses RNAi. We investigated the influence of photolabile protection groups at these and the adjacent nucleobases on siRNA activity and chose to incorporate caged deoxynucleotides instead of ribonucleotides. The siRNAs designed by these means were shown to be completely inactive. By irradiation with UV light (366 nm) they could be fully reactivated and showed the same activity as their unmodified siRNA counterparts.
Mikat
Vera
V
University of Bonn, LIMES-Life and Medical Sciences, Kekulé-Institute, 53121 Bonn, Germany.
Heckel
Alexander
A
D020868:Gene Silencing; D017930:Genes, Reporter; D006367:HeLa Cells / Q000528:radiation effects; D006801:Humans; D008027:Light; D009843:Oligoribonucleotides / Q000737:chemistry; D034622:RNA Interference / Q000528:radiation effects*; D034741:RNA, Small Interfering / Q000235:genetics* / Q000528:radiation effects; D014158:Transcription, Genetic; D014162:Transfection
D009843:Oligoribonucleotides; D034741:RNA, Small Interfering