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G protein–coupled receptors (GPCRs) play a central role in physiological processes and are common drug targets. This GPCR precrystallization screen uses small culture volumes to determine which conditions result in maximal protein expression and stability.
DNA has the capacity to store large amounts of information for very long durations. This protocol describes encoding of digital files as DNA and the error-free retrieval of the stored data from the sequenced data.
Bacterial extracellular vesicles (BEVs) in human body fluids are analyzed using ultrafiltration, size-exclusion chromatography and density-gradient centrifugation to separate the BEVs, followed by post-separation characterization with orthogonal biochemical methods.
Transparent organs are obtained, with retained fluorescent protein signals, upon clearing by immersion in the appropriate CUBIC reagent. The positions of all the cells can be determined using the described software.
This protocol covers the preparation and use of artificial microRNA clusters for gene therapy. The in silico design of chimeric microRNA transgenes in which up to six natural microRNA hairpins are replaced by synthetic versions obviates the need for complex molecular cloning.
hPSCs are differentiated into anterior foregut endoderm and then undergo lineage specification into NKX2-1+ lung progenitor cells. Next, the progenitors undergo distal/alveolar differentiation to produce self-renewing alveolar epithelial type II cells.
This protocol describes how to redesign, characterize, validate and apply genetically encoded dopamine sensors, such as those of the dLight1 family, to measure dopamine transients in cultured cells, neurons, acute brain slices and freely behaving, awake mice.
ICeChIP (internally calibrated ChIP) uses spiked-in, defined nucleosomal standards to overcome the pitfalls of traditional ChIP experiments, enabling the measurement of antibody specificity and the absolute measurement of histone modification density at genomic loci.
Popularization of super-resolution imaging techniques has allowed cell biologists to probe cell structure and function in previously unattainable detail. These methodologies continue to evolve, with new improvements that allow tailoring the available techniques to a particular need and application. This collection showcases primary research articles, reviews and protocols and highlights these recent developments by exemplifying the new, interesting applications of super-resolution microscopy as well as related tool development.