Novel insights into RNAi off-target effects using C. elegans paralogs.
BACKGROUND
In the few years since its discovery, RNAi has turned into a very powerful tool for the study of gene function by allowing post-transcriptional gene silencing. The RNAi mechanism, which is based on the introduction of a double-stranded RNA (dsRNA) trigger whose sequence is similar to that of the targeted messenger RNA (mRNA), is subject to off-target cross-reaction.


RESULTS
We use a novel strategy based on phenotypic analysis of paralogs and predict that, in Caenorhabditis elegans, off-target effects occur when an mRNA sequence shares more than 95% identity over 40 nucleotides with the dsRNA. Interestingly, our results suggest that the minimum length necessary of a high-similarity stretch between a dsRNA and its target in order to observe an efficient RNAi effect varies from 30 to 50 nucleotides rather than 22 nucleotides, which is the length of siRNAs in C. elegans.


CONCLUSIONS
Our predictive methods would improve the design of dsRNA and ultimately the use of RNAi as a therapeutic tool upon experimental verification.
Rual
Jean-François
JF
Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute and Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA. JRUAL@PARTNERS.ORG
Klitgord
Niels
N
Achaz
Guillaume
G
D000818:Animals; D020137:Base Pair Mismatch / Q000235:genetics*; D017173:Caenorhabditis elegans / Q000235:genetics; D029742:Caenorhabditis elegans Proteins / Q000502:physiology; D019295:Computational Biology; D005821:Genetic Techniques / Q000706:statistics & numerical data; D008956:Models, Chemical; D034622:RNA Interference / Q000187:drug effects; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000235:genetics; D043244:Ribonuclease III; D012680:Sensitivity and Specificity
D029742:Caenorhabditis elegans Proteins; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D043244:Ribonuclease III