Specific degradation of 3??? regions of GUS mRNA in posttranscriptionally silenced tobacco lines may be related to 5???-3??? spreading of silencing

Braunstein, Thomas Hartig; Moury, Benoît; Johannessen, M.M.; Albrechtsen, Merete

doi:10.1017/s1355838202026080

Cited by 34 publications

(39 citation statements)

References 37 publications

(23 reference statements)

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“…However, we believe that 3# to 5# spreading over longer distances cannot be excluded because detection of secondary siRNAs was performed 1 week after PVX-U inoculation, thus leaving the possibility that, after a longer period, siRNAs could have spread further along the gus mRNA into the G region. Moreover, the G region might not allow transitivity because it has been shown to be a weak target (English et al, 1996;Braunstein et al, 2002) and it does not give rise to primary siRNAs (Hutvagner et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…In this organism, spreading of silencing is not limited to transgenic targets, but endogenous transcripts can also be used as template for amplification (Sijen et al, 2001;Alder et al, 2003). In plants, transitive silencing can occur in both the 3# to 5# and the 5# to 3# direction along transgenic target RNAs in a primer-dependent or primer-independent manner (Braunstein et al, 2002;Vaistij et al, 2002;Himber et al, 2003;Van Houdt et al, 2003;Kościań ska et al, 2005;Miki et al, 2005;Petersen and Albrechtsen, 2005). Virus-induced gene silencing has been shown to spread over a distance of at least 1,000 nt from the 5# end to the 3# end of the target mRNA, while 3# to 5# spreading can extend at least through 332 nt, with a possible limit of 600 nt (Vaistij et al, 2002;Petersen and Albrechtsen, 2005).…”

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confidence: 99%

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The Frequency and Efficiency of Endogene Suppression by Transitive Silencing Signals Is Influenced by the Length of Sequence Homology

et al. 2006

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Transitivity, the spread of RNA silencing along primary target sequences, leads to the degradation of secondary targets that have no sequence homology to the initial silencing trigger. We demonstrate that increasing the distance between direct and adjacent target sequences in a transgenic primary target delays the onset of silencing of a secondary target gene. Silencing can spread in a 3# to 5# direction over a distance of at least 500 nucleotides (nt), but this requires consistently more time compared to a distance of 98 nt or 250 nt. The efficiency and frequency of transitive silencing of an endogene depends on the length of its sequence homology with the primary target. With a length of 500 nt, efficient silencing can eventually be established in all plants, whereas lengths of 250 nt and 98 nt homology result in less efficient and less frequent suppression. These results suggest that amplification of secondary small interfering RNAs (siRNAs) is a time-requiring process that gradually expands the population of siRNAs until a steady-state level is reached. Moreover, the length of the sequence homology in the primary target providing secondary siRNAs determines whether this steady-state level readily exceeds the threshold necessary for efficient silencing.

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Section: Discussionmentioning

confidence: 99%

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The Frequency and Efficiency of Endogene Suppression by Transitive Silencing Signals Is Influenced by the Length of Sequence Homology

et al. 2006

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“…There is also some indication that RNA silencing can expand across target RNAs and to regions downstream of the primary genome target [20,57,187]. Since transitive RNAi has been reported in plant, invertebrate and vertebrate systems, it is likely that this phenomenon is more widespread than what has been reported but the lack of studies in this area has hampered our knowledge on how widespread the phenomenon of transitive RNAi is.…”

Section: Transgenic Rnaimentioning

confidence: 99%

RNA Interference with Special Reference to Combating Viruses of Crustacea

Fauce

Owens

2012

Indian J. Virol.

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RNA interference has evolved from being a nuisance biological phenomenon to a valuable research tool to determine gene function and as a therapeutic agent. Since pioneering observations regarding RNA interference were first reported in the 1990s from the nematode worm, plants and Drosophila, the RNAi phenomenon has since been reported in all eukaryotic organisms investigated from protozoans, plants, arthropods, fish and mammals. The design of RNAi therapeutics has progressed rapidly to designing dsRNA that can specifically and effectively silence disease related genes. Such technology has demonstrated the effective use of short interfering as therapeutics. In the absence of a B cell lineage in arthropods, and hence no long term vaccination strategy being available, the introduction of using RNA interference in crustacea may serve as an effective control and preventative measure for viral diseases for application in aquaculture.

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“…These newly synthesized siRNAs can target other RNAs on the basis of sequence similarity. In plants, it has been shown that siRNA spreads not only in the 3# to 5# direction but also in the 5# to 3# direction when the green fluorescent protein (gfp) or b-glucuronidase (gus) transgenes are targets of RNA silencing (Braunstein et al, 2002;Klahre et al, 2002;Vaistij et al, 2002;Himber et al, 2003;Van Houdt et al, 2003;Garcia-Perez et al, 2004). However, other reports have shown a lack of such transitive RNA silencing for transgenes.…”

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confidence: 99%

RNA Silencing of Single and Multiple Members in a Gene Family of Rice

2005

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RNA silencing with inverted repeat (IR) constructs has been used to suppress gene expression in various organisms. However, the transitive RNA-silencing effect described in plants may preclude the use of RNA silencing for a gene family. Here, we show that, in rice (Oryza sativa), transitive RNA silencing (spreading of double-stranded RNA along the target mRNA) occurred with the green fluorescent protein transgene but not with the endogenous phytoene desaturase gene. We fused IR copies of unique 3# untranslated regions derived from the rice OsRac gene family to a strong promoter and stably introduced them into rice. Each of the seven members of the OsRac gene family was specifically suppressed by its respective IR construct. We also examined IR constructs in which multiple 3# untranslated regions were fused and showed that three members of the OsRac gene family were effectively suppressed by a single construct. Using highly conserved regions of the two members of the OsRac gene family, we also suppressed the expression of all members of the gene family with variable efficiencies. These results suggest that RNA silencing is a useful method for the functional analysis of gene families in rice and other plants.

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Specific degradation of 3??? regions of GUS mRNA in posttranscriptionally silenced tobacco lines may be related to 5???-3??? spreading of silencing

Abstract: ABSTRACT

Cited by 34 publications

References 37 publications

The Frequency and Efficiency of Endogene Suppression by Transitive Silencing Signals Is Influenced by the Length of Sequence Homology

The Frequency and Efficiency of Endogene Suppression by Transitive Silencing Signals Is Influenced by the Length of Sequence Homology

RNA Interference with Special Reference to Combating Viruses of Crustacea

RNA Silencing of Single and Multiple Members in a Gene Family of Rice

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