Brief Communications in 2010

Context-dependent assembly (CoDA) of zinc finger nucleases is reported. Starting from an archive of zinc finger modules known to function well together, effective multifinger arrays can be constructed using standard techniques. Also in this issue, Doyon et al. report rational design of nucleases with improved cleavage activity.

A statistical framework for assigning confidence scores for protein-protein interaction data generated via affinity purification–mass spectrometry, called significance analysis of interactome (SAINT) is described.

Tracking the displacement of fluorescent beads surrounding a cell embedded in a hydrogel matrix allows quantitative measurement of the three-dimensional traction forces exerted by the cell.

A soft X-ray microscope design using partially incoherent light and a sample holder that can be tilted permits three-dimensional ultrastructural imaging of cryopreserved adherent mammalian cells without chemical fixation.

Protein modules that dimerize rapidly upon exposure to light are reported. They permit light-induced control of dimerization of fused protein targets and can be manipulated with two-photon illumination for experiments in thick samples and in vivo.

A polished and reinforced thinned-skull procedure is used to create a large, chronically stable window in the skull that allows repeated imaging of cortical structures as well as optically guided physiological manipulation. It is proposed as an alternative to the craniotomy and current thinned-skull methods.

An efficient one-step method for re-engineering mouse mutant alleles harboring loxP and FRT sites is reported. It may be applied to the large collection of targeted alleles from the International Knockout Mouse Consortium.

By pooling barcoded genomes of thirty rats before enrichment of a 1.4-megabase target sequence, mutation discovery in 770 genes is achieved with high accuracy.

Magnetic torque tweezers are used to directly measure the torsional stiffness of single molecules of bare DNA and RecA filaments.

Recombinant SV40 viral vectors intravenously injected into mice pretreated with mannitol effectively deliver transgenes to adult neurons in several regions of the central nervous system.

Using 600 oligonucleotides with 60 bases each and three enzymes, the authors assemble the entire mouse mitochondrial genome in four isothermal reactions.

The Trans-ABySS pipeline is an integrated approach for transcript assembly and analysis to identify new mRNA isoforms and structures.

Stimulation of the light-activated cation channel channelrhodopsin-2 can depolarize heart muscle in vitro and in vivo, resulting in precise localized stimulation and constant prolonged depolarization of genetically targeted cardiomyocytes and cardiac tissue.

Adaptations to total internal reflection microscopy permit visualization of the 'footprint' of rolling cells. Applied to neutrophils rolling in whole blood at physiological levels of shear stress, this approach reveals previously unappreciated features of rolling cell biology.

Mice handled by their tails show high levels of anxiety and stress compared to mice handled in cupped hands or in a transparent tunnel.

Random and targeted mutagenesis of the far-red fluorescent protein Katushka followed by screening for low toxicity and red-shifted emission resulted in two near-infrared fluorescent proteins, eqFP650 and eqFP670, that display desirable properties for in vivo imaging compared to existing near-infrared fluorescent proteins.

A gene conferring neomycin resistance can be used for antibiotic selection in C. elegans and C. briggsae. This will permit easy maintenance of transgenic lines and facilitate single-copy insertion of transgenes. Also in this issue, a related paper reports nematode selection using puromycin.

A gene conferring puromycin resistance can be used for antibiotic selection in C. elegans and C. briggsae. This will permit easy maintenance of transgenic lines and facilitate single-copy insertion of transgenes. Also in this issue, a related paper reports nematode selection using neomycin.

Use of a trimethoprim chemical tag allows super-resolution live-cell microscopy by stochastic single molecule–based localization imaging of the dynamics of genetically tagged histone H2B in cell nuclei.

Engineering of the Ca²⁺-sensing domain in existing yellow Cameleon Ca²⁺ indicators is used to create indicators with a range of increased Ca²⁺ affinities capable of detecting subtle changes in intracellular Ca²⁺ at low resting levels.