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Comment
. 2003 May 27;100(11):6289-91.
doi: 10.1073/pnas.1232238100. Epub 2003 May 16.

The specifics of small interfering RNA specificity

Andrew Dillin  1
Affiliations
Comment

The specifics of small interfering RNA specificity

Andrew Dillin. Proc Natl Acad Sci U S A. .
No abstract available

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Figures

Fig. 1.
Fig. 1.
Overview of dsRNA-mediated mRNA degradation. On entry into cells, dsRNA is cleaved by Dicer into 21- to 23-nt siRNAs. siRNAs are complexed with a large multiprotein complex, the RISC. RISC is thought to unwind the siRNA to help target the appropriate mRNA (shown in green). The siRNA/mRNA hybrid is cleaved, releasing the siRNA, and the mRNA is degraded by endo and exonucleases. In worms and plants, the siRNA can also serve as a template for RdRP using the mRNA as a template. Elongation of the siRNA can lead to the production of more siRNAs that could share homology to other genes (shown in orange), causing their degradation, known as transitive RNAi.
Fig. 2.
Fig. 2.
Formal test of transitive RNAi in worms and human 293 cells. (A) In worms, Sijen et al. (13) used GFP–unc-22 fusion genes to test whether mRNA from the endogenous unc-22 gene (shown in red) could be degraded. When dsRNA directed toward GFP was used, the endogenous unc-22 could be degraded only when unc-22 was fused 5′ to the GFP DNA sequences. Fusion at the 3′ end did not cause degradation of the endogenous unc-22 mRNA. (B) Chi et al. (16) used a similar approach to create human kidney cells (293 cells) expressing gene fusions of actin–GFP and actin–luciferase. siRNA directed against GFP caused degradation of the actin–GFP fusion but did not lead to elongation of the siRNA into the actin sequences and subsequent degradation of the actin–luciferase gene fusion. Two siRNAs were used, E1 and E3. E1 is within 20 bp of the fusion junction.
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