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Predictable Transitive RNA Interference Induced by mRNA hairpins in C. elegans
Author(s) -
O'Bier Jennifer,
Cox Jessica,
Doty Matthew,
Schwartz Caitlin,
Rawn J. David,
Dwyer Timothy M.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.787.2
Subject(s) - rna interference , gene silencing , biology , gene , rna silencing , genetics , computational biology , caenorhabditis elegans , trans acting sirna , small hairpin rna , rna
RNA interference (RNAi), is one of the newest and yet most prevalent areas of research today. While the protein machinery necessary to cause this phenomenon has been well characterized, it is not always possible to predict the silencing of one specific gene. At times, when a target gene is silenced, it can lead to the silencing of other RNAs, known as off‐target effects. We are trying to understand this process, termed transitive RNAi, and trying to determine if there are scientific rules governing it. Through the use of an algorithm, we have mapped gene networks that may cause these off‐target effects. One of these networks is now being examined in the nematode C. elegans. Two plasmid constructs have been created. The first (37hp) targets four genes within the network for silencing through a common hairpin sequence found in each of the genes. The second (37a) construct targets just one these genes through sequence homology in an area outside of the hairpin. The two constructs, as well as a control construct, have been introduced into bacteria, induced to express the dsRNA and fed to the C. elgans to induce RNAi. C. elegans fed the bacteria containing the two constructs show slight phenotypic variations. These phenotypic variations lead us to believe that the pattern of silenced genes is similar between the two groups and that the silencing of the one gene with the second construct might have led to the silencing of the other three genes in a manner that we could then begin to predict. The effect of 37a and 37hp constructs against genes within the network are being analyzed, using Real‐time PCR. Preliminary results suggest that the constructs can silence Y37.e3.13 but do not affect Ankyrin. Characterization of the other genes will also be discussed.