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mRNA - Articles and news items
Whitepapers / 18 January 2016 / Breton Hornblower, Ph.D., G. Brett Robb, Ph.D. and George Tzertzinis, Ph.D., New England Biolabs, Inc.
This article discusses options for selection of reagents and the extent to which they influence synthesised mRNA functionality…
Influenza viruses are members of the Orthomyxoviridae family and are a major cause of respiratory tract disease in humans and many animal species. There are three influenza virus types that cause human disease: A, B and C. Type A are further subtyped based on the antigenicity of the hemagglutinin (HA) and neuraminidase (NA) surface proteins and have been associated with serious epidemics and pandemics. Sixteen HA and nine NA subtypes have been identified in birds (one of the natural hosts) and several subtypes have been transmitted to humans. The influenza virus genome is comprised of eight segments of negative-strand RNA that encode for 11 proteins. The genome is under continuous evolution due to random errors in viral replication that allow the virus to evade the host immune system through the genetic processes of antigenic drift and shift. Antigenic drift is the gradual evolution of viral strains due to frequent point mutations within the dominant antibody-binding sites in the HA and NA proteins, which potentially occurs every time the virus replicates. This process drives the need for continuous surveillance of the circulating seasonal influenza strains and their inclusion in the annual vaccination campaign. In contrast, antigenic shift is only seen in influenza A viruses and occurs by re-assortment (i.e., packaging of genes from two or more different viruses). This leads to a new virus subtype, which can result in a pandemic strain if the virus is pathogenic, can spread from human to human and a substantial portion of the population has no preexisting immunity. During the past 100 years, there have been four pandemics (see Table 1 on page 22), three of which have been associated with substantial disease burden and mortality…
Biomedical research often involves the use of cell lines that can be cultured in a laboratory. Individual cells within such cell lines often share a similar morphology. A remarkable exception are in vitro cultured mouse Embryonic Stem Cells (mESCs) – pluripotent cells derived from the blastocyst stage of the mouse developing embryo. Different from many other cell lines, mESCs show heterogeneity in morphology and gene expression between the individual cells within a population. This heterogeneity makes it challenging to characterise mESCs at a molecular level, since global profiling methods generally require large numbers of cells. However, new methods and technologies allow global transcriptome profiling of individual cells. By capturing the global transcriptome of many individual cells using single-cell next-generation mRNA-sequencing, our research focuses on the identification of novel transcription factor modules that contribute to the unique pluripotent state of mESCs, as well as to the differentiation of mESCs, as a model for early mouse embryonic development…
microRNAs (miRNA) are a class of non-coding RNA that regulate the precise amounts of proteins expressed in a cell at a given time. These molecules were discovered in worms in 1993 and only known to exist in humans in the last decade. Despite the youth of the miRNA field, miRNA misexpression is known to occur in a range of human disease conditions and drugs based on modulating miRNA expression are now in development for treatment of cancer, cardiovascular, metabolic and inflammatory diseases. In the last six years, an increasing number of reports have also illuminated diverse roles of cellular miRNAs in viral infection and a miRNA-targeting therapy is currently in phase II clinical trials for treatment of the Hepatitis C virus. Here we review the literature related to miRNAs that regulate viral replication and highlight the factors that will influence the use of miRNA manipulation as a broader antiviral therapeutic strategy.
microRNAs (miRNA) are a class of small noncoding RNA that bind to messenger RNAs (mRNA) and regulate the amount of specific proteins that get expressed. These small RNAs are derived from longer primary transcripts that fold back on themselves to produce stem-loop structures which are recognised and processed by Drosha and co-factors in the nucleus followed by Dicer and co-factors in the cytoplasm, resulting in a ~ 22 nucleotide (nt) duplex RNA, for review see1,2. One strand of the duplex is preferentially incorporated into the RNA-induced silencing complex (RISC) where it then mediates binding to target mRNAs. These interactions lead to decreased protein getting produced from the transcript, due to RNA destabilisation and/or inhibited translation3 (Figure 1). miRNA-mRNA recognition generally requires perfect complementarity with only the first 6-8 nt of a miRNA, termed the ‘seed’ site4. Each miRNA therefore has the potential to interact with hundreds of target mRNAs3,4 and the majority of human protein-coding genes contain miRNA binding sites under selective pressure5. Therapeutic interest in miRNAs has been supported by studies in model organisms demonstrating key functions of individual miRNAs in cancer, cardiac disease, metabolic disease, neuronal and immune cell function6.
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