• Title/Summary/Keyword: post-transcriptional gene silencing

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Characterization of a Tomato (Lycopersicon esculentum Mill.) Ripening-associated Membrane Protein (TRAMP) Gene Expression and Flavour Volatile Changes in TRAMP Transgenic Plants

  • Kim Seog-Hyung;Ji Hee-Chung;Lim Ki-Byung
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.87-95
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    • 2005
  • The tomato ripening associated membrane protein (TRAMP) (Fray et al., 1994) is a member of the major intrinsic protein (MIP) family, defined as channels facilitating the passage of water and small solutes through membranes. During normal fruit ripening the TRAMP mRNA levels were increased whereas the expression levels of TRAMP in low ethylene ACO1-sense suppressed lines, Nr and rin fruits, were lower than at the breaker stage of wild type fruit. TRAMP mRNA is inhibited by $LaCl_3$, which is an inhibitor of $Ca^{2+}$-stimulated responses, treatment but drought condition did not affect TRAMP expression. The levels of TRAMP mRNA transcripts were substantially higher in the dark treated seedlings and fruits. These suggest that TRAMP function as a water channel may be doubted because of several reasons; no water content was changed during ripening in wild type, antisense and overexpression lines, TRAMP expression under light condition was lower than dark condition and TRAMP expression was not changed in drought condition. Co-suppression plant, 3588 was one of sense suppression lines, which contain CaMV 35S promoter and sense pNY507 cDNA, produced small antisense RNA, approximately 21-25 nucleotides in length, mediated post-transcriptional gene silencing. Therefore, TRAMP expression was inhibited by small antisense and multiple copies might induce gene silencing without any production of double strand RNA. Total seven selected volatile productions, isobutylthiazole, 6-methyl-5-hepten-2-one, hexanal, hexenal methylbutanal, hexenol, and methylbutanol, were highly reduced in sense line whereas total volatile production was increased in TRAMP antisense line. These results suggested TRAMP might change volatile related compounds.

Insect Pest Control Technique Using dsRNA (dsRNA를 이용한 해충방제 기술)

  • Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.56 no.2
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    • pp.153-164
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    • 2017
  • Gene silencing using double-stranded RNA (dsRNA) has been widely used in functional genomics in biological organisms. Its principle stems from RNA interference (RNAi), a post-transcriptional control of gene expression. Suppression of specific gene expression using dsRNA may give significant lethal effect. Insect pest control exploits this molecular process to develop novel insecticides using specific dsRNAs. This review explains core principles of RNAi using dsRNA. Then it illustrates various examples to control insect pests using dsRNAs. It also discusses limitations to control insect pests using dsRNAs. Finally, it provides several breakthroughs to develop dsRNA insecticides.

MicroRNA Target Recognition: Insights from Transcriptome-Wide Non-Canonical Interactions

  • Seok, Heeyoung;Ham, Juyoung;Jang, Eun-Sook;Chi, Sung Wook
    • Molecules and Cells
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    • v.39 no.5
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    • pp.375-381
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    • 2016
  • MicroRNAs (miRNAs) are small non-coding RNAs (~22 nucleotides) regulating gene expression at the post-transcriptional level. By directing the RNA-induced silencing complex (RISC) to bind specific target mRNAs, miRNA can repress target genes and affect various biological phenotypes. Functional miRNA target recognition is known to majorly attribute specificity to consecutive pairing with seed region (position 2-8) of miRNA. Recent advances in a transcriptome-wide method of mapping miRNA binding sites (Ago HITS-CLIP) elucidated that a large portion of miRNA-target interactions in vivo are mediated not only through the canonical "seed sites" but also via non-canonical sites (~15-80%), setting the stage to expand and determine their properties. Here we focus on recent findings from transcriptome-wide non-canonical miRNA-target interactions, specifically regarding "nucleation bulges" and "seed-like motifs". We also discuss insights from Ago HITS-CLIP data alongside structural and biochemical studies, which highlight putative mechanisms of miRNA target recognition, and the biological significance of these non-canonical sites mediating marginal repression.