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61.
Christina Falschlehner Sandra Steinbrink Gerrit Erdmann Michael Boutros Professor 《Biotechnology journal》2010,5(4):368-376
RNA interference (RNAi) has become a powerful tool to dissect cellular pathways and characterize gene functions. The availability of genome-wide RNAi libraries for various model organisms and mammalian cells has enabled high-throughput RNAi screenings. These RNAi screens successfully identified key components that had previously been missed in classical forward genetic screening approaches and allowed the assessment of combined loss-of-function phenotypes. Crucially, the quality of RNAi screening results depends on quantitative assays and the choice of the right biological context. In this review, we provide an overview on the design and application of high-throughput RNAi screens as well as data analysis and candidate validation strategies. 相似文献
62.
Therapeutic monoclonal antibodies have revolutionized the treatment of various inflammatory diseases. Immunogenicity against
these antibodies has been shown to be clinically important: it is associated with shorter response duration because of diminishing
concentrations in the blood and with infusion reactions. Concomitant immunomodulators in the form of methotrexate or azathioprine
reduced the immunogenicity of therapeutic antibodies in rheumatoid arthritis, Crohn disease, and juvenile idiopathic arthritis.
The occurrence of adverse events does not increase when immunomodulators are added to therapeutic antibodies. The mechanism
whereby methotrexate and azathioprine influence immunogenicity remains unclear. Evidence-based consensus on prescribing concomitant
immunomodulators is needed. 相似文献
63.
Corradi N Croll D Colard A Kuhn G Ehinger M Sanders IR 《Applied and environmental microbiology》2007,73(1):366-369
Gene copy number polymorphism was studied in a population of the arbuscular mycorrhizal fungus Glomus intraradices by using a quantitative PCR approach on four different genomic regions. Variation in gene copy number was found for a pseudogene and for three ribosomal genes, providing conclusive evidence for a widespread occurrence of macromutational events in the population. 相似文献
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Room temperature crystal structure of the fast switching M159T mutant of the fluorescent protein dronpa
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Marius Kaucikas Ann Fitzpatrick Elana Bryan Abelone Struve Robert Henning Irina Kosheleva Vukica Srajer Gerrit Groenhof Jasper J. Van Thor 《Proteins》2015,83(3):397-402
The fluorescent protein Dronpa undergoes reversible photoswitching reactions between the bright “on” and dark “off” states via photoisomerization and proton transfer reactions. We report the room temperature crystal structure of the fast switching Met159Thr mutant of Dronpa at 2.0‐Å resolution in the bright on state. Structural differences with the wild type include shifted backbone positions of strand β8 containing Thr159 as well as an altered A‐C dimer interface involving strands β7, β8, β10, and β11. The Met159Thr mutation increases the cavity volume for the p‐hydroxybenzylidene‐imidazolinone chromophore as a result of both the side chain difference and the backbone positional differences. Proteins 2015; 83:397–402. © 2014 Wiley Periodicals, Inc. 相似文献
67.
Bioprocessing analysis of Pyrococcus furiosus strains engineered for CO2‐based 3‐hydroxypropionate production
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68.
Mark Barnes Gerrit van Rensburg Wai-Ming Li Kashif Mehmood Sebastian Mackedenski Ching-Man Chan Dustin T. King Andrew L. Miller Chow H. Lee 《The Journal of biological chemistry》2015,290(1):625-639
The ability of its four heterogeneous nuclear RNP-K-homology (KH) domains to physically associate with oncogenic mRNAs is a major criterion for the function of the coding region determinant-binding protein (CRD-BP). However, the particular RNA-binding role of each of the KH domains remains largely unresolved. Here, we mutated the first glycine to an aspartate in the universally conserved GXXG motif of the KH domain as an approach to investigate their role. Our results show that mutation of a single GXXG motif generally had no effect on binding, but the mutation in any two KH domains, with the exception of the combination of KH3 and KH4 domains, completely abrogated RNA binding in vitro and significantly retarded granule formation in zebrafish embryos, suggesting that any combination of at least two KH domains cooperate in tandem to bind RNA efficiently. Interestingly, we found that any single point mutation in one of the four KH domains significantly impacted CRD-BP binding to mRNAs in HeLa cells, suggesting that the dynamics of the CRD-BP-mRNA interaction vary over time in vivo. Furthermore, our results suggest that different mRNAs bind preferentially to distinct CRD-BP KH domains. The novel insights revealed in this study have important implications on the understanding of the oncogenic mechanism of CRD-BP as well as in the future design of inhibitors against CRD-BP function. 相似文献
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