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1.
Sci Rep ; 13(1): 3235, 2023 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-36828850

RESUMO

Naturally abundant dyes are very attractive for the development of dye-sensitized solar cells (DSSCs). Hydroxycinnamic acid derivatives, such as caffeic acid (CA), ferulic acid (FA), and p-coumaric acid (PA), were considered for the selective harvesting of ultraviolet A (UVA) (315-400 nm) photons. Their spectroscopic and electrochemical properties were investigated both theoretically and experimentally. They were further successfully adopted as photosensitizers in UV-selective and visibly transparent DSSCs, which exhibited a power conversion efficiency of 0.22-0.38% under AM (air mass) 1.5G (global) illumination (100 mW/cm2) and 3.40-3.62% under UVA irradiation (365 nm, 115.22 mW/cm2), with a corresponding visible light transmittance (VLT) of 49.07-43.72% and a general color rendering index (Ra) of 93-90.


Assuntos
Corantes , Energia Solar , Corantes/química , Ácidos Cumáricos , Fontes de Energia Elétrica , Raios Ultravioleta
2.
Chem Asian J ; 18(6): e202201142, 2023 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-36710260

RESUMO

A combustion-assisted polyol reduction (CPR) method has been developed to deposit electrocatalytically efficient and transparent Pt counter electrodes (CEs) for bifacial dye-sensitized solar cells (DSSCs). Compared with conventional thermal decomposition of Pt precursors, CPR allows for a decrease in reduction temperature to 150 °C. The low-temperature processing is attributed to adding an organic fuel, acetylacetone (Hacac), which provides extra heat to lower reduction energy. In addition, the stable Pt complexes can simultaneously be formed in ethylene glycol (EG) and Hacac system, which leads to Pt nanoparticle size regulation. A ratio of Hacac to EG is optimized to achieve excellent electrocatalytic activity and high visible light transmittance for CEs. The bifacial DSSCs fabricated with CPR-Pt CEs (EG : Hacac=1 : 16) reach efficiencies of 6.71±0.16% and 6.41±0.15% in front and back irradiations, respectively.

3.
Colloids Surf A Physicochem Eng Asp ; 640: 128418, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35125661

RESUMO

The significant public health concerns related to particulate matter (PM) air pollutants and the airborne transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have led to considerable interest in high-performance air filtration membranes. Highly ferroelectric polyvinylidene fluoride (PVDF) nanofiber (NF) filter membranes are successfully fabricated via electrospinning for high-performance low-cost air filtration. Spectroscopic and ferro-/piezoelectric analyses of PVDF NF show that a thinner PVDF NF typically forms a ferroelectric ß phase with a confinement effect. A 70-nm PVDF NF membrane exhibits the highest fraction of ß phase (87%) and the largest polarization behavior from piezoresponse force microscopy. An ultrathin 70-nm PVDF NF membrane exhibits a high PM0.3 filtration efficiency of 97.40% with a low pressure drop of 51 Pa at an air flow of 5.3 cm/s owing to the synergetic combination of the slip effect and ferroelectric dipole interaction. Additionally, the 70-nm PVDF NF membrane shows excellent thermal and chemical stabilities with negligible filtration performance degradation (air filtration efficiency of 95.99% and 87.90% and pressure drop of 55 and 65 Pa, respectively) after 24 h of heating at 120 °C and 1 h immersion in isopropanol.

4.
Nanomaterials (Basel) ; 10(3)2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32168882

RESUMO

Pt catalytic nanoparticles on F-doped SnO2/glass substrates were prepared by polyol reduction below 200 °C. The polyol reduction resulted in better transparency of the counter electrode and high power-conversion efficiency (PCE) of the resultant dye-sensitized solar cells (DSSCs) compared to conventional thermal reduction. The PCEs of the DSSCs with 5 µm-thick TiO2 photoanodes were 6.55% and 5.01% under front and back illumination conditions, respectively. The back/front efficiency ratio is very promising for efficient bifacial DSSCs.

5.
ACS Appl Mater Interfaces ; 9(29): 24763-24770, 2017 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-28670900

RESUMO

We report a new p-type dopant, manganese oxide (Mn3O4) nanoparticle, to enhance the performance of organic field-effect transistors (OFETs) with conjugated polymers, including poly(3-hexylthiophene-2,5-diyl), poly[[N,N 9-bis(2-octyldodecyl)-naphthalene-1,4,5,8-bis(dicarboximide)-2,6-diyl]-alt-5,59-(2,29-bithiophene)], and poly[[2,5-bis(2-octyldodecyl)-2,3,5,6-tetrahydro-3,6-dioxopyrrolo[3,4-c]pyrrole-1,4-diyl]-alt-[[2,2'-(2,5-thiophene)bis-thieno(3,2b) thiophene]-5,5'-diyl]] (DPPT-TT). Incorporating a small amount of Mn3O4 nanoparticles in the semiconductor film significantly improved the hole mobility and decreased the threshold voltage for all OFETs, indicating efficient Mn3O4 nanoparticle p-type doping. The Mn3O4 nanoparticle showed a better doping efficiency than the widely used FeCl3 dopant due to better mixability with the host conjugated polymers. In particular, doped DPPT-TT OFETs showed significantly improved mobility up to 2.35 (±0.4) cm2/(V·s) with enhanced air and operational stability at 0.1 wt % doping concentration from 1.2 cm2/(V·s) for pristine devices.

6.
Phys Chem Chem Phys ; 18(34): 23904-9, 2016 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-27523163

RESUMO

The high performance of soluble [60]fulleropyrrolidine upon its use as the active layer of n-channel organic field-effect transistors (OFETs) is reported. The two materials, N-phenyl derivatives C60-fused-N-phenyl-2-phenylpyrrolidine ([C60]PhNPh) and C60-fused N-phenyl-2-hexylpyrrolidine ([C60]HexNPh), have well-controlled molecular structures with a modification of the pyrrolidine ring, with no increase in the LUMO level, achieving a high mobility and highly ambient stable n-type OFET. The top-gate, bottom-contact device shows a high electron charge-carrier mobility of up to 0.14 and 0.08 cm(2) V(-1) s(-1) for [C60]PhNPh and [C60]HexNPh, respectively, (Ion/Ioff = 10(6)) with the commonly used CYTOP dielectric. Excess carriers introduced by a small amount of chemical doping of polyethyleneimine (PEI) compensate traps by shifting the Fermi level (EF) toward the respective transport energy levels and therefore increase charge-carrier mobility (0.26 and 0.1 cm(2) V(-1) s(-1)) and provide good ambient operational stability compared with pristine devices.

7.
Sci Rep ; 6: 29811, 2016 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-27440253

RESUMO

The general form of interfacial contact resistance was derived for organic thin-film transistors (OTFTs) covering various injection mechanisms. Devices with a broad range of materials for contacts, semiconductors, and dielectrics were investigated and the charge injections in staggered OTFTs was found to universally follow the proposed form in the diffusion-limited case, which is signified by the mobility-dependent injection at the metal-semiconductor interfaces. Hence, real ohmic contact can hardly ever be achieved in OTFTs with low carrier concentrations and mobility, and the injection mechanisms include thermionic emission, diffusion, and surface recombination. The non-ohmic injection in OTFTs is manifested by the generally observed hook shape of the output conductance as a function of the drain field. The combined theoretical and experimental results show that interfacial contact resistance generally decreases with carrier mobility, and the injection current is probably determined by the surface recombination rate, which can be promoted by bulk-doping, contact modifications with charge injection layers and dopant layers, and dielectric engineering with high-k dielectric materials.

8.
RNA Biol ; 13(6): 554-60, 2016 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-27144897

RESUMO

microRNAs (miRNAs) are an abundant class of small endogenous non-coding RNAs (ncRNAs) of ∼22 nucleotides (nts) in length. These small regulatory molecules are involved in diverse developmental, physiological and pathological processes. miRNAs target mRNAs (mRNAs) for translational repression and/or mRNA degradation. Predictions of miRNA binding sites facilitate experimental validation of miRNA targets. Models developed with data from CLIP studies have been used for predictions of miRNA binding sites in the whole transcriptomes of human, mouse and worm. The prediction results have been assembled into STarMirDB, a new database of miRNA binding sites available at http://sfold.wadsworth.org/starmirDB.php . STarMirDB can be searched by miRNAs or mRNAs separately or in combination. The search results are categorized into seed and seedless sites in 3' UTR, CDS and 5' UTR. For each predicted site, STarMirDB provides a comprehensive list of sequence, thermodynamic and target structural features that are known to influence miRNA: target interaction. A high resolution PDF diagram of the conformation of the miRNA:target hybrid is also available for visualization and publication. The results of a database search are available through both an interactive viewer and downloadable text files.


Assuntos
Caenorhabditis elegans/genética , Bases de Dados de Ácidos Nucleicos , MicroRNAs/genética , RNA Mensageiro/genética , Regiões 3' não Traduzidas , Regiões 5' não Traduzidas , Animais , Sítios de Ligação , Regulação da Expressão Gênica , Humanos , Camundongos , Estabilidade de RNA , RNA Mensageiro/química , Análise de Sequência de RNA , Software , Interface Usuário-Computador
9.
Phys Chem Chem Phys ; 18(18): 12486-93, 2016 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-27087486

RESUMO

To determine the role played by the choice of processing solvents in governing the photophysics, microstructure, and charge carrier transport in naphthalenediimide (NDI)-based polymers, we have prepared two new NDI-bithiophene (T2)- and NDI-thienothiophene (TTh)-containing polymers with hybrid siloxane pentyl chains (SiC5) (P(NDI2SiC5-T2) and P(NDI2SiC5-TTh)). Among the various processing solvents studied here, the films prepared using chloroform exhibited far better electron mobilities (0.16 ± 0.1-0.21 ± 0.05 cm(2) V(-1) s(-1)) than the corresponding samples prepared from different solvents, exceeding one order of magnitude higher, indicating the significant influence of the processing solvent on the charge transport. Upon thin-film analysis using atomic force microscopy and grazing incidence X-ray diffraction, we discovered that molecular ordering and orientation are affected by the choice of the processing solvent, which is responsible for the change in the transport characteristics of this class of polymers.

10.
Phys Chem Chem Phys ; 17(31): 20160-7, 2015 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-26179975

RESUMO

A simply and facilely synthesized MoO3 solution was developed to fabricate charge injection layers for improving the charge-injection properties in p-type organic field-effect transistors (OFETs). By dissolving MoO3 powder in ammonium (NH3) solvent under an air atmosphere, an intermediate ammonium molybdate ((NH4)2MoO4) precursor is made stable, transparent and spin-coated to form the MoO3 interfacial layers, the thickness and morphology of which can be well-controlled. When the MoO3 layer was applied to OFETs with a cost-effective molybdenum (Mo) electrode, the field-effect mobility (µFET) was significantly improved to 0.17 or 1.85 cm(2) V(-1)s(-1) for polymer semiconductors, regioregular poly(3-hexylthiophene) (P3HT) or 3,6-bis-(5bromo-thiophen-2-yl)-N,N'-bis(2-octyl-1-dodecyl)-1,4-dioxo-pyrrolo[3,4-c]pyrrole (DPPT-TT), respectively. Device analysis indicates that the MoO3-deposited Mo contact exhibits a contact resistance RC of 1.2 MΩ cm comparable to that in a device with the noble Au electrode. Kelvin-probe measurements show that the work function of the Mo electrode did not exhibit a dependence on the thickness of MoO3 film. Instead, ultraviolet photoemission spectroscopy results show that a doping effect is probably induced by casting the MoO3 layer on the P3HT semiconductor, which leads to the improved hole injection.

11.
Nucleic Acids Res ; 42(Web Server issue): W114-8, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24803672

RESUMO

STarMir web server predicts microRNA (miRNA) binding sites on a target ribonucleic acid (RNA). STarMir is an implementation of logistic prediction models developed with miRNA binding data from crosslinking immunoprecipitation (CLIP) studies (Liu,C., Mallick, B., Long, D., Rennie, W.A., Wolenc, A., Carmack, C.S. and Ding, Y. (2013). CLIP-based prediction of mammalian microRNA binding sites. Nucleic Acids Res., 41(14), e138). In both intra-dataset and inter-dataset validations, the models showed major improvements over established algorithms in predictions of both seed and seedless sites. General applicability of the models was indicated by good performance in cross-species validations. The input data for STarMir is processed by the web server to perform prediction of miRNA binding sites, compute comprehensive sequence, thermodynamic and target structure features and a logistic probability as a measure of confidence for each predicted site. For each of seed and seedless sites and for all three regions of a mRNA (3' UTR, CDS and 5' UTR), STarMir output includes the computed binding site features, the logistic probability and a publication-quality diagram of the predicted miRNA:target hybrid. The prediction results are available through both an interactive viewer and downloadable text files. As an application module of the Sfold RNA package (http://sfold.wadsworth.org), STarMir is freely available to all at http://sfold.wadsworth.org/starmir.html.


Assuntos
MicroRNAs/metabolismo , RNA Mensageiro/metabolismo , Software , Animais , Sítios de Ligação , Humanos , Internet , Camundongos , MicroRNAs/química , RNA Mensageiro/química , Análise de Sequência de RNA
12.
RNA Biol ; 11(6): 693-701, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24827614

RESUMO

MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression. Since the discovery of lin-4, the founding member of the miRNA family, over 360 miRNAs have been identified for Caenorhabditis elegans (C. elegans). Prediction and validation of targets are essential for elucidation of regulatory functions of these miRNAs. For C. elegans, crosslinking immunoprecipitation (CLIP) has been successfully performed for the identification of target mRNA sequences bound by Argonaute protein ALG-1. In addition, reliable annotation of the 3' untranslated regions (3' UTRs) as well as developmental stage-specific expression profiles for both miRNAs and 3' UTR isoforms are available. By utilizing these data, we developed statistical models and bioinformatics tools for both transcriptome-scale and developmental stage-specific predictions of miRNA binding sites in C. elegans 3' UTRs. In performance evaluation via cross validation on the ALG-1 CLIP data, the models were found to offer major improvements over established algorithms for predicting both seed sites and seedless sites. In particular, our top-ranked predictions have a substantially higher true positive rate, suggesting a much higher likelihood of positive experimental validation. A gene ontology analysis of stage-specific predictions suggests that miRNAs are involved in dynamic regulation of biological functions during C. elegans development. In particular, miRNAs preferentially target genes related to development, cell cycle, trafficking, and cell signaling processes. A database for both transcriptome-scale and stage-specific predictions and software for implementing the prediction models are available through the Sfold web server at http://sfold.wadsworth.org.


Assuntos
Regiões 3' não Traduzidas , Sítios de Ligação , Caenorhabditis elegans/genética , MicroRNAs/genética , RNA Mensageiro/genética , Animais , Pareamento de Bases , Sequência de Bases , Caenorhabditis elegans/embriologia , Biologia Computacional/métodos , Bases de Dados de Ácidos Nucleicos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Masculino , MicroRNAs/química , RNA Mensageiro/química , Curva ROC , Software , Transcriptoma
13.
PLoS One ; 8(10): e75149, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24204572

RESUMO

Microsatellites or simple sequence repeats (SSRs) are one of the most popular sources of genetic markers and play a significant role in plant genetics and breeding. In this study, we identified citrus SSRs in the genome of Clementine mandarin and analyzed their frequency and distribution in different genomic regions. A total of 80,708 SSRs were detected in the genome with an overall density of 268 SSRs/Mb. While di-nucleotide repeats were the most frequent microsatellites in genomic DNA sequence, tetra-nucleotides, which had more repeat units than any other SSR types, had the highest cumulative sequence length. We identified 6,834 transcripts as containing 8,989 SSRs in 33,929 Clementine mandarin transcripts, among which, tri-nucleotide motifs (36.0%) were the most common, followed by di-nucleotide (26.9%) and hexa-nucleotide motifs (15.1%). The motif AG (16.7%) was most abundant among these SSRs, while motifs AAG (6.6%), AAT (5.0%), and TAG (2.2%) were most common among tri-nucleotides. Functional categorization of transcripts containing SSRs revealed that 5,879 (86.0%) of such transcripts had homology with known proteins, GO and KEGG annotation revealed that transcripts containing SSRs were those implicated in diverse biological processes in plants, including binding, development, transcription, and protein degradation. When 27 genomic and 78 randomly selected SSRs were tested on Clementine mandarin, 95 SSRs revealed polymorphism. These 95 SSRs were further deployed on 18 genotypes of the three generas of Rutaceae for the genetic diversity assessment, genomic SSRs generally show low transferability in comparison to SSRs developed from expressed sequences. These transcript-markers identified in our study may provide a valuable genetic and genomic tool for further genetic research and varietal development in citrus, such as diversity study, QTL mapping, molecular breeding, comparative mapping and other genetic analyses.


Assuntos
Genômica , Repetições de Microssatélites/genética , Citrus/classificação , Citrus/genética , Genoma de Planta , Estudo de Associação Genômica Ampla , Anotação de Sequência Molecular , Motivos de Nucleotídeos , Fases de Leitura Aberta , Filogenia , Polimorfismo Genético , Especificidade da Espécie , Transcriptoma , Regiões não Traduzidas
14.
Nucleic Acids Res ; 41(14): e138, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23703212

RESUMO

Prediction and validation of microRNA (miRNA) targets are essential for understanding functions of miRNAs in gene regulation. Crosslinking immunoprecipitation (CLIP) allows direct identification of a huge number of Argonaute-bound target sequences that contain miRNA binding sites. By analysing data from CLIP studies, we identified a comprehensive list of sequence, thermodynamic and target structure features that are essential for target binding by miRNAs in the 3' untranslated region (3' UTR), coding sequence (CDS) region and 5' untranslated region (5' UTR) of target messenger RNA (mRNA). The total energy of miRNA:target hybridization, a measure of target structural accessibility, is the only essential feature common for both seed and seedless sites in all three target regions. Furthermore, evolutionary conservation is an important discriminating feature for both seed and seedless sites. These features enabled us to develop novel statistical models for the predictions of both seed sites and broad classes of seedless sites. Through both intra-dataset validation and inter-dataset validation, our approach showed major improvements over established algorithms for predicting seed sites and a class of seedless sites. Furthermore, we observed good performance from cross-species validation, suggesting that our prediction framework can be valuable for broad application to other mammalian species and beyond. Transcriptome-wide binding site predictions enabled by our approach will greatly complement the available CLIP data, which only cover small fractions of transcriptomes and known miRNAs due to non-detectable levels of expression. Software and database tools based on the prediction models have been developed and are available through Sfold web server at http://sfold.wadsworth.org.


Assuntos
MicroRNAs/metabolismo , RNA Mensageiro/química , Regiões 3' não Traduzidas , Regiões 5' não Traduzidas , Algoritmos , Proteínas Argonautas/metabolismo , Sítios de Ligação , Bases de Dados de Ácidos Nucleicos , Células HEK293 , Humanos , Imunoprecipitação/métodos , Modelos Logísticos , RNA Mensageiro/metabolismo , Software
15.
Development ; 136(9): 1497-507, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19336466

RESUMO

It is important to understand the regulation of stem cell division because defects in this process can cause altered tissue homeostasis or cancer. The cyclin-dependent kinase inhibitor Dacapo (Dap), a p21/p27 homolog, acts downstream of the microRNA (miRNA) pathway to regulate the cell cycle in Drosophila melanogaster germline stem cells (GSCs). Tissue-extrinsic signals, including insulin, also regulate cell division of GSCs. We report that intrinsic and extrinsic regulators intersect in GSC division control; the Insulin receptor (InR) pathway regulates Dap levels through miRNAs, thereby controlling GSC division. Using GFP-dap 3'UTR sensors in vivo, we show that in GSCs the dap 3'UTR is responsive to Dicer-1, an RNA endonuclease III required for miRNA processing. Furthermore, the dap 3'UTR can be directly targeted by miR-7, miR-278 and miR-309 in luciferase assays. Consistent with this, miR-278 and miR-7 mutant GSCs are partially defective in GSC division and show abnormal cell cycle marker expression, respectively. These data suggest that the GSC cell cycle is regulated via the dap 3'UTR by multiple miRNAs. Furthermore, the GFP-dap 3'UTR sensors respond to InR but not to TGF-beta signaling, suggesting that InR signaling utilizes Dap for GSC cell cycle regulation. We further demonstrate that the miRNA-based Dap regulation may act downstream of InR signaling; Dcr-1 and Dap are required for nutrition-dependent cell cycle regulation in GSCs and reduction of dap partially rescues the cell cycle defect of InR-deficient GSCs. These data suggest that miRNA- and Dap-based cell cycle regulation in GSCs can be controlled by InR signaling.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Células Germinativas/metabolismo , Proteínas Nucleares/metabolismo , RNA Helicases/metabolismo , Receptor de Insulina/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismo , Regiões 3' não Traduzidas/genética , Animais , Animais Geneticamente Modificados , Divisão Celular , Proteínas de Drosophila/genética , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Regulação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Células Germinativas/citologia , MicroRNAs/genética , Proteínas Nucleares/genética , RNA Helicases/genética , Receptor de Insulina/genética , Ribonuclease III , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo
16.
BMC Bioinformatics ; 10 Suppl 1: S33, 2009 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-19208134

RESUMO

BACKGROUND: RNA interference (RNAi) mediated by small interfering RNAs (siRNAs) or short hairpin RNAs (shRNAs) has become a powerful technique for eukaryotic gene knockdown. siRNA GC-content negatively correlates with RNAi efficiency, and it is of interest to have a convincing mechanistic interpretation of this observation. We here examine this issue by considering the secondary structures for both the target messenger RNA (mRNA) and the siRNA guide strand. RESULTS: By analyzing a unique homogeneous data set of 101 shRNAs targeted to 100 endogenous human genes, we find that: 1) target site accessibility is more important than GC-content for efficient RNAi; 2) there is an appreciable negative correlation between GC-content and RNAi activity; 3) for the predicted structure of the siRNA guide strand, there is a lack of correlation between RNAi activity and either the stability or the number of free dangling nucleotides at an end of the structure; 4) there is a high correlation between target site accessibility and GC-content. For a set of representative structural RNAs, the GC content of 62.6% for paired bases is significantly higher than the GC content of 38.7% for unpaired bases. Thus, for a structured RNA, a region with higher GC content is likely to have more stable secondary structure. Furthermore, by partial correlation analysis, the correlation for GC-content is almost completely diminished, when the effect of target accessibility is controlled. CONCLUSION: These findings provide a target-structure-based interpretation and mechanistic insight for the effect of GC-content on RNAi efficiency.


Assuntos
Interferência de RNA , RNA Mensageiro/química , RNA Interferente Pequeno/química , Composição de Bases , Citosina/análise , Guanina/análise , Conformação de Ácido Nucleico , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , RNA não Traduzido/química , RNA não Traduzido/genética
17.
Mol Ther ; 17(3): 538-47, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19088704

RESUMO

RNA interference (RNAi) can be an effective antiviral agent; however, overexpression of RNAi can be toxic through competition with the endogenous microRNA (miRNA) machinery. We used rational design to identify highly potent RNAi that is effective at nontoxic doses. A statistical analysis was conducted to pinpoint thermodynamic characteristics correlated with activity. Sequences were selected that conformed to a consensus internal stability profile (ISP) associated with active RNAi, and RNAi triggers were expressed in the context of an endogenous miRNA. These approaches yielded highly active hepatitis B virus (HBV) RNAi. A statistical analysis found a correlation between activity and nucleation by binding within the seed sequence to accessible regions in the target RNA. Guide strands were selected for favorable strand biasing, but increased strand biasing did not correlate with potency, suggesting a threshold effect. Exogenous short hairpin RNAs (shRNAs), but not miRNAs were previously reported to compete with miRNAs for the miRNA/RNAi machinery. In contrast, we show that exogenous Polymerase III- but not Polymerase II-driven miRNAs compete with exogenous miRNAs, at multiple steps in the miRNA pathway. Exogenous miRNAs also compete with endogenous miR-21. Thus, competition with endogenous miRNAs should be monitored even when using miRNA-based therapeutics. However, potent silencing was achieved at doses where competition was not observed.


Assuntos
Vírus da Hepatite B/genética , Interferência de RNA , Sequência de Bases , Linhagem Celular Tumoral , Humanos , MicroRNAs/química , MicroRNAs/genética , Dados de Sequência Molecular , Conformação de Ácido Nucleico , RNA Mensageiro/genética , Termodinâmica , Transcrição Gênica/genética
18.
Pac Symp Biocomput ; : 64-74, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18232104

RESUMO

MicroRNAs (miRNAs) are small non-coding RNAs that repress protein synthesis by binding to target messenger RNAs (mRNAs) in multicellular eukaryotes. The mechanism by which animal miRNAs specifically recognize their targets is not well understood. We recently developed a model for modeling the interaction between a miRNA and a target as a two-step hybridization reaction: nucleation at an accessible target site, followed by hybrid elongation to disrupt local target secondary structure and form the complete miRNA-target duplex. Nucleation potential and hybridization energy are two key energetic characteristics of the model. In this model, the role of target secondary structure on the efficacy of repression by miRNAs is considered, by employing the Sfold program to address the likelihood of a population of structures that co-exist in dynamic equilibrium for a specific mRNA molecule. This model can accurately account for the sensitivity to repression by let-7 of both published and rationally designed mutant forms of the Caenorhabditis elegans lin-41 3' UTR, and for the behavior of many other experimentally-tested miRNA-target interactions in C. elegans and Drosophila melanogaster. The model is particularly effective in accounting for certain false positive predictions obtained by other methods. In this study, we employed this model to analyze a set of miRNA-target interactions that were experimentally tested in mammalian models. These include targets for both mammalian miRNAs and viral miRNAs, and a viral target of a human miRNA. We found that our model can well account for both positive interactions and negative interactions. The model provides a unique explanation for the lack of function of a conserved seed site in the 3' UTR of the viral target, and predicts a strong interaction that cannot be predicted by conservation-based methods. Thus, the findings from this analysis and the previous analysis suggest that target structural accessibility is generally important for miRNA function in a broad class of eukaryotic systems. The model can be combined with other algorithms to improve the specificity of predictions by these algorithms. Because the model does not involve sequence conservation, it is readily applicable to target identification for microRNAs that lack conserved sites, non-conserved human miRNAs, and poorly conserved viral mRNAs. StarMir is a new Sfold application module developed for the implementation of the structure-based model, and is available through Sfold Web server at http://sfold.wadsworth.org.


Assuntos
MicroRNAs/genética , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Animais , Sítios de Ligação , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Biologia Computacional , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Hepacivirus/genética , Hepacivirus/metabolismo , Mamíferos , MicroRNAs/química , Modelos Genéticos , Modelos Moleculares , Conformação de Ácido Nucleico , Hibridização de Ácido Nucleico , RNA de Helmintos/genética , RNA de Helmintos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Viral/genética , RNA Viral/metabolismo , Termodinâmica
19.
Nat Methods ; 5(9): 813-9, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19160516

RESUMO

Target prediction for animal microRNAs (miRNAs) has been hindered by the small number of verified targets available to evaluate the accuracy of predicted miRNA-target interactions. Recently, a dataset of 3,404 miRNA-associated mRNA transcripts was identified by immunoprecipitation of the RNA-induced silencing complex components AIN-1 and AIN-2. Our analysis of this AIN-IP dataset revealed enrichment for defining characteristics of functional miRNA-target interactions, including structural accessibility of target sequences, total free energy of miRNA-target hybridization and topology of base-pairing to the 5' seed region of the miRNA. We used these enriched characteristics as the basis for a quantitative miRNA target prediction method, miRNA targets by weighting immunoprecipitation-enriched parameters (mirWIP), which optimizes sensitivity to verified miRNA-target interactions and specificity to the AIN-IP dataset. MirWIP can be used to capture all known conserved miRNA-mRNA target relationships in Caenorhabditis elegans at a lower false-positive rate than can the current standard methods.


Assuntos
Caenorhabditis elegans/genética , MicroRNAs/genética , RNA Mensageiro/genética , Ribonucleoproteínas/genética , Animais , Sítios de Ligação , Proteínas de Transporte/metabolismo , MicroRNAs/metabolismo
20.
Nat Struct Mol Biol ; 14(4): 287-94, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17401373

RESUMO

MicroRNAs (miRNAs) are small noncoding RNAs that repress protein synthesis by binding to target messenger RNAs. We investigated the effect of target secondary structure on the efficacy of repression by miRNAs. Using structures predicted by the Sfold program, we model the interaction between an miRNA and a target as a two-step hybridization reaction: nucleation at an accessible target site followed by hybrid elongation to disrupt local target secondary structure and form the complete miRNA-target duplex. This model accurately accounts for the sensitivity to repression by let-7 of various mutant forms of the Caenorhabditis elegans lin-41 3' untranslated region and for other experimentally tested miRNA-target interactions in C. elegans and Drosophila melanogaster. These findings indicate a potent effect of target structure on target recognition by miRNAs and establish a structure-based framework for genome-wide identification of animal miRNA targets.


Assuntos
MicroRNAs/metabolismo , Conformação de Ácido Nucleico , RNA de Helmintos/química , Regiões 3' não Traduzidas/metabolismo , Animais , Sequência de Bases , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/metabolismo , Drosophila melanogaster/metabolismo , Dados de Sequência Molecular , Proteínas Mutantes/metabolismo , Hibridização de Ácido Nucleico , Nucleotídeos/metabolismo , RNA de Helmintos/genética , Análise de Regressão , Software , Relação Estrutura-Atividade , Termodinâmica , Fatores de Transcrição/metabolismo
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