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A proximity assay based on methylation of interacting 'prey' proteins by a 'bait' fused to the histone lysine methyltransferase permits the detection of enzyme-substrate protein-protein interactions in yeast.
Structure determination followed by targeted engineering of the popular photoactivatable fluorescent protein monomeric (m)Eos2 yields mEos3 versions that are more monomeric and less disruptive in protein fusions and also exhibit higher labeling density, brightness and other beneficial properties.
Structured illumination using multifocal patterned illumination via a digital micromirror device integrated into a conventional wide-field microscope, followed by digital processing, allows resolution-doubled three-dimensional imaging of live organisms with two-color capability.
A robot, algorithm and software for automated in vivo intracellular electrophysiology are reported that can automatically perform whole-cell patch clamping in the living mouse brain with quality comparable to that for a trained human experimenter.
Simultaneous functional magnetic resonance imaging (fMRI) and fiber-optic–based calcium recordings in rats allow investigation of the relationship between blood oxygen level–dependent (BOLD) fMRI signals and the underlying neural activity. The study uncovers prolonged BOLD signal components involving glial activation.
A hybrid fluorescence molecular tomography–X ray computed tomography system is applied for in vivo imaging of multiple mouse models, and its performance is validated on post-mortem cryosection data.
Nanobodies that bind to fluorescent proteins with high affinity and are coupled to bright organic dyes allow simple efficient labeling of fusion proteins for super-resolution microscopy.
Global optimization of single-molecule localizations using compressed sensing allows stochastic optical reconstruction microscopy (STORM) at high molecular densities and live cell super-resolution imaging with a temporal resolution of 3 seconds.
The freely available WormToolbox enables high-throughput image analysis of a variety of phenotypes of Caenorhabditis elegans in liquid culture and should prove useful for image-based screens.
The authors analyze how sequencing depth, choice of control sample, paired-end versus single-end reads and the selection of peak-calling algorithm influence the interpretation of chromatin immunoprecipitation–sequencing (ChIP-seq) experiments.
Microbial Assemblage Prediction is a predictive model for the climate-dependent abundance of microbial taxa in space and time. It takes potential interactions between taxa into account and is used on longitudinal metagenomic and climate data from the Western English Channel.
To increase the efficiency of direct neuronal conversion of postnatal human fibroblasts, the authors combine two-factor neuronal programming with small molecules. This method increases the yield and purity of functional neuron-like cells by more than 15-fold.
The use of marker coselection in combination with multiplex automated genome engineering (MAGE) is reported to improve the efficiency of engineered changes in bacterial genomes. The authors use the method to insert twelve 20-base-pair T7 promoters to control indirubin and indigo production.
Reprogramming of mouse fibroblasts to induced pluripotency is demonstrated in suspension culture. Also in this issue, Fluri et al. report suspension-culture reprogramming of mouse cells and further differentiation, also in suspension, into cardiac cells.
The authors compare segregation of a protein into two daughter cells for the wild-type protein and a fluorescently tagged version, by assessing protein activity in the two cells; differences in segregation between the two protein versions indicate mislocalization artifacts caused by the fluorescent tag. Using this system they identify widespread artifacts in the localization of bacterial proteases.
A computational framework is reported for the accurate and sensitive identification of RNA editing sites from whole-genome DNA and RNA sequences from the same individual.
Transgenic expression of secreted antibodies specific for modified heparan sulfates fused to GFP allow the visualization of these modifications in vivo.
