Use of cell cultures in vitro to assess the uptake of long dsRNA in plant cells

In Vitro Cellular & Developmental Biology - Plant - While RNAi approaches based on transgenic plants overexpressing double-stranded RNAs (dsRNAs) have yielded a number of genetically modified...     

Lateral hydrological connectivity differentially affects the community characteristics of multiple groups of aquatic invertebrates in tropical wetland pans in South Africa

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Livestock disturbances in Mediterranean temporary ponds: A mesocosm experiment with sheep manure and simulated trampling

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The double-stranded RNA-binding protein,Staufen1, is an IRES-transacting factor regulating HIV-1 cap-independent translation initiation

Translation initiation of the viral genomic mRNA (vRNA) of human immunodeficiency virus-type 1 (HIV-1) can be mediated by a cap- or an internal ribosome entry site (IRES)-dependent mechanism. A previous report shows that Staufen1, a cellular double-stranded (ds) RNA-binding protein (RBP), binds to the 5’untranslated region (5′UTR) of the HIV-1 vRNA and promotes its cap-dependent translation. In this study, we now evaluate the role of Staufen1 as an HIV-1 IRES-transacting factor (ITAF). We first confirm that Staufen1 associates with both the HIV-1 vRNA and the Gag protein during HIV-1 replication. We found that in HIV-1-expressing cells, siRNA-mediated depletion of Staufen1 reduces HIV-1 vRNA translation. Using dual-luciferase bicistronic mRNAs, we show that the siRNA-mediated depletion and cDNA-mediated overexpression of Staufen1 acutely regulates HIV-1 IRES activity. Furthermore, we show that Staufen1-vRNA interaction is required for the enhancement of HIV-1 IRES activity. Interestingly, we find that only Staufen1 harboring an intact dsRNA-binding domain 3 (dsRBD3) rescues HIV-1 IRES activity in Staufen1 CRISPR-Cas9 gene edited cells. Finally, we show that the expression of Staufen1-dsRBD3 alone enhances HIV-1 IRES activity. This study provides evidence of a novel role for Staufen1 as an ITAF promoting HIV-1 vRNA IRES activity.     

RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) AND PARSIMONY METHODS

Abstract — Random amplified polymorphic DNA (RAPD) data possess a number of undesirable features for parsimony analysis. These features include their non-codominant inheritance, their anonymous nature, their different (a)symmetrical transformation probabilities, and their possible GC priming bias. As a consequence, no single parsimony method seems appropriate for RAPD data. Moreover, the presence/absence coding of RAPDs is equivalent to the invalid independent allele model for allozymes. These issues are discussed and the way in which parsimony analysis of RAPDs can be misleading is illustrated.