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Independent two-color, two-photon uncaging of glutamate and GABA allows autonomous activation and inhibition of neuronal action potentials in brain slices with subcellular resolution.
A chronically implanted biocompatible electrochemical microsensor allows long-term recording of subsecond dopamine dynamics in vivo. The microsensor can reliably detect behaviorally evoked dopamine release from dopamine neurons in the brain over a period of months in rats.
A mouse strain in which cellular reprogramming factors are expressed from a defined genomic locus is presented. It will enable studies of reprogramming in multiple cell types as well as facilitate comparisons between induced pluripotent stem cells and embryonic stem cells. Also in this issue, a paper by Carey et al. presents related tools.
Mouse strains in which three or four cellular reprogramming factors are expressed from a defined genomic locus are presented. They will enable studies of reprogramming in multiple cell types as well as facilitate comparisons between induced pluripotent stem cells and embryonic stem cells. Also in this issue, a paper from Stadtfeld et al. presents related tools.
An efficient system for the reversion and modification of mouse gene trap alleles is presented. It is applicable to available collections of gene trap embryonic stem cell lines.
Selected reaction monitoring (SRM) is a powerful mass spectrometry technology to reliably detect selected protein targets, even those at very low abundance, but requires tedious assay development for each protein of interest. High-throughput SRM assay development is now possible by using crude synthetic peptide libraries without purification to represent each protein target.
Single-molecule sequencing of poly(A)-tailed chromatin immunoprecipitated DNA proves equal in sensitivity and accuracy to amplification-based sequencing technologies and allows analysis of samples sizes as small as 50 pg.
Glycan structure, attachment site and the glycoprotein from which it came can be identified with a method to enrich for glycoproteins from complex biological samples, digest them on a bead and release the glycopeptides for mass spectrometry analysis.
A cocktail of three small molecules improves the efficiency of reprogramming human fibroblasts to induced pluripotent stem cells and allows survival of the cells after trypsinization.
Activity of yeast cytosine deaminase can be both positively and negatively selected by adjusting growth conditions. Adapting this life-death selection to a protein complementation assay based on the enzyme allows dissection of protein-protein interactions and protein functions in yeast.
Quantitative information is necessary to determine which protein interactions are the most relevant in a cellular context. A defined set of affinity purification experiments combined with quantitative mass spectrometry analysis allows the determination of dissociation constants for all protein targets interacting with an introduced ligand.
Using an axial detector, scanning transmission electron microscopy allows three-dimensional tomographic reconstruction of micrometer-thick sections of biological samples, at a resolution comparable to that obtained on thin sections.
Technical modifications of existing methods lead to a somatic cell nuclear transfer method in the zebrafish, which yields adult cloned fish with healthy offspring that carry donor traits.
A series of genetically encoded fluorescent sensors for intracellular Zn2+ with binding affinities spanning the picomolar and nanomolar ranges show that cytosolic Zn2+ is buffered at ∼0.4 nM. Targeting of the sensors to insulin-containing secretory granules indicates a free Zn2+ concentration between 1 and 100 μM in these organelles.
A human embryonic stem cell line, ErythrRED, harbors a red fluorescent protein under the control of regulatory sequences from the beta-globin locus, as a reporter for erythroid differentiation.
A general approach to address the 'phase problem' in protein crystallography is described, allowing protein structures to be directly solved from 2 Å resolution diffraction data without using heavy atom doping or relying on a preexisting structure model for molecular replacement.
Deletions of polyguanine tracts in Caenorhabditis elegans deficient in the DOG-1 DNA helicase can be exploited to generate deletion alleles of several genes for which no such alleles exist in this organism.
PyroNoise, an algorithm that preclusters the flowgrams generated on a 454 GS FLX with DNA extracted from microbial samples can distinguish between noise and genuine sequence diversity in a metagenomic dataset.
This array with 623,124 SNP probes and 916,269 probes to query structural variants opens the door to a detailed characterization of genetic diversity in laboratory mouse strains. This will allow genome-wide association studies in mice.
To deplete abundant rRNA from total RNA during cDNA library generation, hexamer primers that perfectly match rRNA are removed. These 'not so randomly primed' cDNA libraries can be generated from small amount of total RNA, preserve strand orientation and are equally enriched in polyadenylated and non-polyadenylated transcripts.
