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An optimized transcription activator–like effector (TALE) and an improved assembly method promise efficient genome editing and transcriptome modulation.
The combination of an ultrahigh-resolution dual optical trap with a confocal microscope allowed single-fluorophore detection of labeled oligonucleotide binding and simultaneous measurement of angstrom-scale changes in DNA tether extension.
Confined photoactivation of photoactivatable mCherry using two-photon illumination with line-scanning temporal focusing in combination with three-dimensional localization algorithms allows three-dimensional super-resolution microscopy of cellular features at <50 nm lateral and <100 nm axial resolution and depths greater than 8 μm.
A microfluidic mixing device for multiple, rapid and automated single-molecule measurements permits the study of macromolecule properties under varying environmental conditions. Also in this issue, Gambin et al. present another microfluidic mixing device for rapid single-molecule measurements.
A laminar flow mixing microfluidic device enables single-molecule fluorescence resonance energy transfer (FRET) kinetic measurements with a time resolution of 0.2 ms, enabling the study of early binding-coupled folding and unfolding events of an intrinsically disordered protein, α-synuclein. Also in this issue, Kim et al. describe another microfluidic mixing device for single-molecule experiments.
A genetic multicolor cell-labeling technique for Droshophila melanogaster, Flybow, is described and applied to the study of neural circuits. This method implements a variant of the mouse Brainbow strategy in combination with specific neuronal targeting using the Gal-4–upstream activating sequence system to select for membrane-tethered fluorescent proteins. Also in this issue, Hampel et al. report a similar strategy, Drosophila Brainbow, to select for epitope-tagged proteins detectable via immunofluorescence.
A genetic multicolor cell-labeling technique for Droshophila melanogaster, Drosophila Brainbow, is described and applied to the study of neural circuits. This method implements a variant of the mouse Brainbow strategy in combination with specific neuronal targeting using the Gal-4–upstream activating sequence system to select for epitope-tagged proteins detectable with immunofluorescence. Also in this issue, Hadjieconomou et al. develop a similar strategy, Flybow, to select for membrane-tethered fluorescent proteins.
Changing the codon sequence in Caenorhabditis elegans genes allows fine-tuning of transgene expression from high to low expression. The same strategy is likely applicable for Drosophila melanogaster and Saccharomyces cerevisiae.
By methylating the phosphate groups of PtdIns(3,4,5)P3 researchers can load this lipid more efficiently into a mass spectrometer and thus this lipid can be quantified in the presence of an internal synthetic standard.
A genetic platform allows refinement of tissue-specific expression using the upstream activating sequence–GAL4 system in Drosophila melanogaster, facilitating the segmentation of complex expression patterns and allowing GAL4 expression patterns to be repurposed.