A versatile, high-efficiency platform for CRISPR-based gene activation

AbstractCRISPR-mediated transcriptional activation (CRISPRa) is a powerful technology for inducing gene expression from endogenous loci with exciting applications in high throughput gain-of-function genomic screens and the engineering of cell-based models. However, current strategies for generating potent, stable, CRISPRa-competent cell lines present limitations for the broad utility of this approach. Here, we provide a high-efficiency, self-selecting CRISPRa enrichment strategy, which combined with piggyBac transposon technology enables rapid production of CRISPRa-ready cell populations compatible with a variety of downstream assays. We complement this with an optimized guide RNA scaffold that significantly enhances CRISPRa functionality. Finally, we describe a synthetic guide RNA tool set that enables transient, population-wide gene activation when used with the self-selecting CRISPRa system. Taken together, this versatile platform greatly enhances the potential for CRISPRa across a wide variety of cellular contexts.</jats:p

Effect of oxygen and biofilms on crevice corrosion of UNS S31803 and UNS N08825 in natural seawater

The effect of oxygen and biofilms on crevice corrosion of UNS S31803 and UNS N08825 in seawater was studied. Passivity breakdown occurred through crevice corrosion in UNS N08825 and through transpassive dissolution in UNS S31803 although both alloys displayed crevice corrosion under potentiodynamic conditions. The most severe crevice corrosion occurred in the absence of oxygen and the presence of a biofilm. Microbial adhesion as investigated by fluorescence microscopy occurred mainly outside the crevice and DNA sequencing revealed a shift in biofilm composition as a function of substratum surface and oxygen pressure

Publisher Correction: Global site-specific neddylation profiling reveals that NEDDylated cofilin regulates actin dynamics (Nature Structural &amp; Molecular Biology, (2020), 27, 2, (210-220), 10.1038/s41594-019-0370-3).

In the version of this article initially published online, in Fig. 6d, the third and fourth bars were incorrectly labeled “DMSO + cytochrome D” and “MLN4924 + cytochrome D,” respectively. They should have been labeled “DMSO + cytochalasin D” and “MLN4924 + cytochalasin D,” respectively. The errors have been corrected in the print, PDF and HTML versions of the article