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Results for “mRNA sequencing”

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At least 163 records · Page 9Linked to original sources
PubMed1998

Differential intracellular sorting of immediate early gene mRNAs depends on signals in the mRNA sequence.

This study characterizes the differential targeting of recently synthesized immediate early gene (IEG) mRNAs to neuronal cell bodies versus dendrites and tests the hypothesis that this targeting is based on signals in the encoded proteins. A single electroconvulsive seizure induces the expression of a number of IEG mRNAs in granule cells of the dentate gyrus. Most of these IEG mRNAs remain in the cell body, including two that are characterized in the present study (the mRNAs for NGFI-A and COX-2). In contrast, the mRNA for Arc moved rapidly into dendrites at an apparent rate of approximately 300 micron/hr. Inhibiting protein synthesis with cycloheximide did not disrupt the differential mRNA sorting, demonstrating that the differential targeting of mRNAs is not dependent on translation.

Animals↗
PubMed1996

Lack of DNase I mRNA sequences in murine lenses.

Clarity of the mammalian lens is due in part to the complete lack of internal organelles, including nuclei, within the lens fiber cells that compose the bulk of the lens. Experimental evidence shows that as the differentiation of lens fiber cells progresses, nuclei and nuclear DNA are actively degraded. Prior characterization of chick lens development suggests that DNase I could be involved in lens DNA degredation. However, recent data suggest that DNase I is unlikely to be the nuclease responsible for DNA degredation in the differentiating lens. In this report, we find that in the murine lens, mRNA for DNase I is undetectable by northern blotting or PCR. We conclude that mRNA for DNase I is either not present or present in very low levels in murine lens. Our results are consistent with the hypothesis that DNase I is not involved in lens DNA degredation.

Animals↗
PubMed2025

Droplet-Based Single-Cell 3' mRNA Sequencing of Marburg Virus-Infected Samples.

Single-cell technologies are continually evolving with emerging methods that are gradually uncovering the central DNA-RNA-protein dogma. Single-cell RNA sequencing is one arm of a multi-omic approach that achieves an astounding level of granularity to reveal the complexity of virus-host interactions at the transcriptomic level. Cell tropism, virus replication, pathogenesis, and gene expression changes mediated by the virus and the host's immune response to infection are just some areas of study that are gaining better clarity due to the high-resolution analysis afforded by the technology.We describe a single-cell sequencing protocol for Marburg virus infection in vivo using nonhuman primate blood and the 10× Chromium Next GEM single-cell genomics methodology. Working with pathogens of high consequence is logistically complicated, requiring containment in biosafety level (BSL)-4 laboratories and harsh inactivation procedures before samples can safely be removed to lower biosafety conditions. We provide procedural insight into sample isolation and processing conducted in BSL-4 and describe the requirements for safe sample removal without jeopardizing quality for down-stream sequencing and analysis in BSL-2 conditions. Characterization of complicated biological processes mediated by high-containment pathogens, typically restricted to analogous model systems, e.g., minigenome, can be achieved using live virus.

Animals↗
PubMed1987

Molecular cloning of mRNA sequences transiently induced during rat liver regeneration.

In order to isolate genes which are induced during liver regeneration, we have constructed a cDNA library from 16-h-regenerating liver poly(A)+ RNA. By computer analysis of autoradiograms produced by differential plaque hybridization with cDNA from normal or 16-h-regenerating liver, we have isolated several recombinant clones representing sequences transiently increased during liver regeneration. Three of these were further characterized: the level of the corresponding mRNAs increase rapidly after partial hepatectomy, before the onset of DNA synthesis. Two clones were identified as fibrinogen clones. It is speculated that alpha-fibrinogen may be involved in the growth process, or in its regulation.

Animals↗
PubMed1995

The 3' untranslated region of bovine preprolactin contains a transferable non-poly(A) mRNA sequence that prolongs translation.

Preprolactin transcripts, synthesized in vitro, were actively translated for a prolonged period when injected into Xenopus oocytes. As a result, prolactin continued to be secreted into the media for up to 6 days after injection of the transcript. To investigate the role of the preprolactin 3' untranslated sequence in stabilizing transcripts, it was fused to coding regions derived from signal recognition particle receptor alpha-subunit or preproinsulin receptor. The translational half-life of the chimeric RNA was increased for both coding regions, suggesting that a sequence within the preprolactin 3' untranslated region that prolongs translation is transferable. Deletion mutagenesis of this untranslated region demonstrated that a sequence of 98 nucleotides immediately following the prolactin stop codon was sufficient to prolong translation of RNAs injected into Xenopus oocytes.

Animals↗