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A strategy, based on solid-state NMR spectroscopy, for determining the structure of an oligomeric, seven-helix membrane protein (Anabaena sensory rhodopsin) in a lipid environment is described.
This work describes wide-field temporal focusing, a two-photon volumetric imaging technique based on light sculpting that enables functional imaging of the majority of neurons in the head ganglia of C. elegans with high temporal and spatial resolution.
Addition of weak helper interactions to fluorescent protein pairs by protein engineering provides a simple method to increase FRET efficiency with little or no background.
CRISPR-Cas9–mediated cleavage is used to stimulate homologous recombination at specific target sites in the C. elegans genome, permitting flexible tagging and sequence modification of endogenous worm genes.
Single DNA-binding proteins can be tracked on densely covered DNA at high spatial and temporal resolution and in the presence of high protein concentrations by using a technique that combines optical tweezers, confocal fluorescence microscopy and stimulated emission depletion (STED) nanoscopy.
An algorithm and software tool to uncover contact networks of interacting conformationally heterogeneous protein residues from X-ray crystallography data is described.
Targeting unique variants in highly identical paralogous genes with molecular inversion probes followed by high-throughput sequencing will open a way to associate features in these duplicated genes with human traits.
The synthetic promoter E-SARE provides a genetic tool to tag neurons in an activity-dependent manner. The authors show the utility of this tool for labeling populations of neurons that respond to specific stimuli in living mice and for tracking the axonal projection patterns.
Identifying tissue-specific expression of a selection marker and putative enhancer-driven expression of GFP with flow cytometry enriches for active enhancers.
A cellular engineering approach coupled with mass spectrometry allows the cell-of-origin of intra- and extracellular proteins to be determined from co-cultured cells.
A method to tightly attach cell-derived extracellular matrix (ECM) to the culture surface is described. It is applied to generate bone marrow mesenchymal stem cell–derived ECM, which supports culture of human hematopoietic stem and progenitor cells.
A semiconductor chip, coupled to an ion-sensitive field effect transistor (ISFET) pH sensor, can amplify and quantitate DNA in real time without dyes, cameras and external heating devices.
Two methods for identifying protein isoforms that are concurrently phosphorylated and ubiquitylated are applied in yeast to identify phosphorylation sites that regulate ubiquitin proteasome–mediated proteome degradation.
A method for enzymatically producing long, high-purity, single-stranded DNA oligonucleotides should find many applications in basic research, in DNA nanotechnology and in clinical fields.
For genetically engineered circuits, the movement of RNA polymerase across the DNA during transcription needs to be tightly controlled. A large library of strong terminators will make circuit design easier and more efficient.
Algorithms that account for and overcome the intrinsic pixel-dependent readout noise from scientific complementary metal-oxide semiconductor (sCMOS) cameras prevent localization artifacts and avoid substantial compromises in localization uncertainty in single-molecule imaging. When combined with multi-emitter fitting, sCMOS cameras allow video-rate single-molecule switching nanoscopy.
Automated methods are described for the determination of survival curves in C. elegans, enabling rapid and statistically rigorous studies of lifespan in this organism.