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Modified dsRNAs that are not processed by Dicer maintain potency and are incorporated into the RISC.

ABSTRACT: Chemical modification of RNA duplexes can provide practical advantages for RNA interference (RNAi) triggering molecules including increased stability, safety and specificity. The impact of nucleotide modifications on Dicer processing, RISC loading and RNAi-mediated mRNA cleavage was investigated with duplexes >or=25 bp in length. It is known that dsRNAs >or=25 bp are processed by Dicer to create classic 19-bp siRNAs with 3'-end overhangs. We demonstrate that the presence of minimal modification configurations on longer RNA duplexes can block Dicer processing and result in the loading of the full-length guide strand into RISC with resultant mRNA cleavage at a defined site. These longer, modified duplexes can be highly potent gene silencers, with EC50s in the picomolar concentration range, demonstrating that Dicer processing is not required for incorporation into RISC or potent target silencing.

SUBMITTER: Salomon W

PROVIDER: S-EPMC2887946 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature

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Modified dsRNAs that are not processed by Dicer maintain potency and are incorporated into the RISC.

Salomon William W Bulock Karen K Lapierre Jennifer J Pavco Pamela P Woolf Tod T Kamens Joanne J

Nucleic acids research 20100218 11

Chemical modification of RNA duplexes can provide practical advantages for RNA interference (RNAi) triggering molecules including increased stability, safety and specificity. The impact of nucleotide modifications on Dicer processing, RISC loading and RNAi-mediated mRNA cleavage was investigated with duplexes >or=25 bp in length. It is known that dsRNAs >or=25 bp are processed by Dicer to create classic 19-bp siRNAs with 3'-end overhangs. We demonstrate that the presence of minimal modification ...[more]

PMID: 20167638

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Project description:Humic acid-enhanced phosphate fertilizer (HAP) is widely applied in Chinese agriculture due to its high efficiency. Although the structural composition and physicochemical properties of humic acid (HA) are significantly altered during HAP production, a clear understanding of the mechanisms underlying the biological effects of HA extracted from HAP fertilizer (PHA) on plant growth is still lacking. In the current study, we extracted PHA from HAP and assessed its effects on the dry biomass, phosphorus (P) and nitrogen (N) uptake, and P absorption rate of maize seedlings when supplied at different concentrations (2.5, 5, 10, and 25 mg C L-1) in the hydroponic culture. The root vigor, root plasma membrane H+-ATPase activity, and root nitrate reductase activity were also determined as the representative indicators of the root capacity for nutrient absorption, and used to clarify the mechanism by which PHA affects the maize growth and nutrient absorption. The results showed that the dry biomass, phosphorus uptake, nitrogen uptake, and average phosphorus absorption rates were significantly higher by 14.7-27.9%, 9.6-35.1%, 17.9-22.4%, and 22.1-31.0%, respectively, in plants treated with 2.5-5 mg C L-1 PHA compared to untreated controls. Application of 10-25 mg C L-1 raw HA resulted in similar stimulatory effects on plant growth and nutrient absorption. However, higher levels of PHA (10-25 mg C L-1) negatively impacted these indicators of plant growth. Furthermore, low PHA or high raw HA concentrations similarly improved root vigor and root plasma membrane H+-ATPase and nitrate reductase (NR) activities. These results indicate that lower concentrations of PHA can stimulate maize seedling growth and nutrient absorption to an extent that is comparable to the effect of higher concentrations of raw HA. Thus, the proportion of HA incorporated into HAP could be lower than the theoretical amount estimated through assays evaluating the biological effects of raw HA.

Dataset Information

Modified dsRNAs that are not processed by Dicer maintain potency and are incorporated into the RISC.

Publications

Modified dsRNAs that are not processed by Dicer maintain potency and are incorporated into the RISC.

Similar Datasets

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