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RNA Interference in Infectious Tropical Diseases

The Korean Journal of Parasitology 2008;46(1):1-15.
Published online: March 20, 2008

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Corresponding author (skang1@tulane.edu)
• Received: January 8, 2008   • Accepted: February 12, 2008

Copyright © 2008 by The Korean Society for Parasitology

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Korean J Parasitol. 2008;46(1):1-15.   Published online March 20, 2008
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RNA Interference in Infectious Tropical Diseases
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Fig. 1 Target mRNA degradation by RNAi gene silencing. Dicer initiates RNAi by cleaving dsRNAs into ~22 bp small interfering RNAs (siRNAs). During RISC assembly, Ago2 directly binds to the siRNA and cleaves sense siRNAs (blue strands) and only the anti-sense siRNAs (red strands) remain associated with the RISC complex. After assembly of RISC, the antisense strand directs RISC to target mRNAs. The RISC cut the target mRNAs at 11 to 12 bp downstream of the 5' end of the antisense strand covering the target mRNA.
Fig. 2 Functions of SID-1 and SID-2 in systemic RNAi in C. elegans. (A) SID-1 as a channel, allowing dsRNAs to diffuse into and between cells. sid-1 mutant C. elegans or sid-1-deficient worms neither can take up dsRNAs from environment nor can spread between cells. Therefore, both microinjection and feeding methods are not effective to deliver dsRNAs. This RNAi-deficiency can be rescued by heterologous expression of C. elegans wild type sid-1 [96,97]. (B) SID-2 also acts as a channel for dsRNAs but is only localized in the apical intestinal lumen. Thus, SID-2 is responsible for dsRNA uptake from environment (e.g. from lumen to pseudocoelom), but not for spread between cells. Feeding dsRNA is not effective for gene silencing in sid-2 mutant C. elegans or sid-2-deficient worms, but microinjection can be used for dsRNA delivery because dsRNAs in the pseudocoelom can be spread systemically via SID-1. sid-2 mutants can be rescued for RNAi by heterologous expression of a wild copy of C. elegans sid-2 [96,98].
Fig. 3 Strategies for generation of dsRNA in vivo by symmetric transcription. (A) SympUAST-w produces dsRNAs of the w gene by simultaneous transcription using two identical UAS promoters flanking the target gene in opposite directions. The target gene is transcribed in both directions and the resulting sense and antisense RNAs are hybridized to form dsRNAs. (B) pUAST-IRsp-w contains inverted repeats of the w gene with a spacer between the repeats. This is a common approach to generate hairpin dsRNAs. (C) pUAST-IR-w contains inverted repeats of the w gene without a spacer. This could generate hairpin dsRNAs, but the dsRNAs were not efficient enough to silence a target gene. This may be due to deletions in the center of inverted repeats, rendering the hairpin structure unstable. it appears that symmetrically transcribed dsRNA system may be effective enough to replace the inverted repeat hairpin RNAi system [113].
RNA Interference in Infectious Tropical Diseases