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2.
Sci Rep ; 10(1): 11139, 2020 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-32636422

RESUMO

Transgenic maize plants expressing dsRNA targeting western corn rootworm (WCR, Diabrotica virgifera virgifera) DvSSJ1 mRNA, a Drosophila snakeskin (ssk) ortholog, show insecticidal activity and significant plant protection from WCR damage. The gene encodes a membrane protein associated with the smooth sepate junction (SSJ) which is required for intestinal barrier function. To understand the active RNA form that leads to the mortality of WCR larvae by DvSSJ1 RNA interference (RNAi), we characterized transgenic plants expressing DvSSJ1 RNA transcripts targeting WCR DvSSJ1 mRNA. The expression of the silencing cassette results in the full-length transcript of 901 nucleotides containing a 210 bp inverted fragment of the DvSSJ1 gene, the formation of a double-stranded RNA (dsRNA) transcript and siRNAs in transgenic plants. Our artificial diet-feeding study indicates that dsRNAs greater than or equal to approximately 60 base-pairs (bp) are required for DvSSJ1 insecticidal activity. Impact of specificity of dsRNA targeting DvSSJ1 mRNA on insecticidal activities was also evaluated in diet bioassay, which showed a single nucleotide mutation can have a significant impact or abolish diet activities against WCR. These results provide insights as to the functional forms of plant-delivered dsRNA for the protection of transgenic maize from WCR feeding damage and information contributing to the risk assessment of transgenic maize expressing insecticidal dsRNA.


Assuntos
Besouros , Controle Biológico de Vetores/métodos , Plantas Geneticamente Modificadas/genética , Zea mays/genética , Animais , Besouros/metabolismo , Proteínas de Insetos/genética , Junções Intercelulares/metabolismo , Larva , Interferência de RNA , RNA de Cadeia Dupla/genética
3.
PeerJ ; 8: e8577, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32110490

RESUMO

Several data loggers deployed to monitor temperature and humidity of Crocodylus acutus (American crocodile) nests in South Florida could not be located after hatching. One badly damaged data logger was retrieved, providing insight into the possible fate of the others. Using a taphonomic approach, we identified numerous indentations, consistent with crocodylian bite marks, and inconsistent with potential mammalian or squamate bites. It seems most likely that the data logger was damaged by the nesting C. acutus rather than during attempted nest predation. Estimated bite forces for reproductive age, female C. acutus exceed the predicted material properties of the data logger's housing, suggesting that the bites were exploratory in nature. We suggest that data loggers be removed prior to hatching or permit remote data storage.

4.
BMC Genomics ; 18(1): 290, 2017 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-28403821

RESUMO

BACKGROUND: Motility and the coordination of moving food through the gastrointestinal tract rely on a complex network of neurons known as the enteric nervous system (ENS). Despite its critical function, many of the molecular mechanisms that direct the development of the ENS and the elaboration of neural network connections remain unknown. The goal of this study was to transcriptionally identify molecular pathways and candidate genes that drive specification, differentiation and the neural circuitry of specific neural progenitors, the phox2b expressing ENS cell lineage, during normal enteric nervous system development. Because ENS development is tightly linked to its environment, the transcriptional landscape of the cellular environment of the intestine was also analyzed. RESULTS: Thousands of zebrafish intestines were manually dissected from a transgenic line expressing green fluorescent protein under the phox2b regulatory elements [Tg(phox2b:EGFP) w37 ]. Fluorescence-activated cell sorting was used to separate GFP-positive phox2b expressing ENS progenitor and derivatives from GFP-negative intestinal cells. RNA-seq was performed to obtain accurate, reproducible transcriptional profiles and the unbiased detection of low level transcripts. Analysis revealed genes and pathways that may function in ENS cell determination, genes that may be identifiers of different ENS subtypes, and genes that define the non-neural cellular microenvironment of the ENS. Differential expression analysis between the two cell populations revealed the expected neuronal nature of the phox2b expressing lineage including the enrichment for genes required for neurogenesis and synaptogenesis, and identified many novel genes not previously associated with ENS development. Pathway analysis pointed to a high level of G-protein coupled pathway activation, and identified novel roles for candidate pathways such as the Nogo/Reticulon axon guidance pathway in ENS development. CONCLUSION: We report the comprehensive gene expression profiles of a lineage-specific population of enteric progenitors, their derivatives, and their microenvironment during normal enteric nervous system development. Our results confirm previously implicated genes and pathways required for ENS development, and also identify scores of novel candidate genes and pathways. Thus, our dataset suggests various potential mechanisms that drive ENS development facilitating characterization and discovery of novel therapeutic strategies to improve gastrointestinal disorders.


Assuntos
Sistema Nervoso Entérico/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Proteínas de Homeodomínio/genética , Análise de Sequência de RNA/métodos , Fatores de Transcrição/genética , Peixe-Zebra/genética , Animais , Diferenciação Celular , Linhagem da Célula , Movimento Celular , Microambiente Celular , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Neurogênese
5.
Int Rev Cell Mol Biol ; 308: 273-321, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24411174

RESUMO

Understanding the process by which an uncommitted dividing cell produces particular specialized cells within a tissue remains a fundamental question in developmental biology. Many tissues are well suited for cell-fate studies, but perhaps none more so than the developing retina. Traditionally, experiments using the retina have been designed to elucidate the influence that individual environmental signals or transcription factors can have on cell-fate decisions. Despite a substantial amount of information gained through these studies, there is still much that we do not yet understand about how cell fate is controlled on a systems level. In addition, new factors such as noncoding RNAs and regulators of chromatin have been shown to play roles in cell-fate determination and with the advent of "omics" technology more factors will most likely be identified. In this chapter we summarize both the traditional view of retinal cell-fate determination and introduce some new ideas that are providing a challenge to the older way of thinking about the acquisition of cell fates.


Assuntos
Diferenciação Celular , Retina/citologia , Retina/metabolismo , Animais , Humanos
6.
Genome Med ; 5(4): 35, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23673197

RESUMO

Personalized genomics will only be useful for monitoring the prognosis of patients with cancer when it becomes much more cost-effective and quicker to apply. A recent study brings this closer to reality with the development of plasma-seq, a rapid, low-cost method that sequences the circulating DNA present in the peripheral blood of patients with cancer. The power of this technique is demonstrated with the examination of tumor genomes from patients with prostate cancer. SEE RELATED RESEARCH ARTICLE: http://genomemedicine.com/content/5/4/30.

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