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Packaging split Cas9 into AAVs increases cargo capacity and allows for efficient genome editing and gene activation in vivo. AAV–split-Cas9 activates the host immune system but does not trigger the extensive cellular damage observed with delivery of Cas9 via DNA electroporation.
The integration of ligand-responsive riboswitches with single guide RNAs allows Cas9–effector fusions to be targeted in response to ligands and thereby translate a cellular signal into a downstream readout of choice.
Flexible mesh electronics facilitate stable long-term recordings of the same single neurons in mouse brains over months, enabling chronic recordings in behaving animals and longitudinal studies to resolve aging-dependent changes in neural activity.
uDISCO clearing renders whole animals transparent and capitalizes on shrinkage to image them. This method allows the analysis of intact nervous systems and whole-body screens for transplanted cells and human tissue samples after prolonged storage.
Synthetic DNA spike-ins that recapitulate genetic variation present in human genomes serve as quantitative and qualitative controls for genome sequencing and variant detection.
Synthetic spike-in standards (‘sequins’), representing spliced mRNA isoforms, provide internal controls for assessing transcript assembly and quantification within and between RNA sequencing libraries. Sequins representing fused genes can be used to determine the sensitivity limit for oncogenic fusions in cancer samples.
A network-based method and computational tool, PIUMet, reveals disease-associated molecular pathways from untargeted metabolomics data without requiring mass-spectral feature identification.
TRIC, a cross-run alignment algorithm and software tool, enables reproducible quantification of thousands of peptides across multiple targeted liquid chromatography–tandem mass spectrometry runs.
A bright and photostable far-red fluorescent protein, smURFP, was developed from a cyanobacterial phycobiliprotein. smURFP uniquely binds a highly cell-permeable biliverdin derivative to obtain fluorescence brightness comparable to that of eGFP in cells.
Reversible cryo-arrest allows consecutive super-resolution and functional imaging of molecular patterns in the same mammalian cell. This method was used to study the evolution of receptor tyrosine kinase reaction patterns at multiple scales.
ExFISH extends expansion microscopy to single-molecule RNA imaging, enabling super-resolution imaging of diverse RNAs in cells and tissues on conventional microscopes. The method enables multiplexed imaging of RNA and improved RNA quantitation.
UMI-4C is a rapid, simplified barcoding approach to targeted chromatin conformation capture that produces high-complexity libraries from low sample input, is easily multiplexed and gives a quantitative, statistically defined readout.
When studying neural circuitry, the ablation of synapses may be an alternative to optogenetic manipulation of neurons. A genetically encoded tool called GFE3 eliminates inhibitory inputs into neurons expressing GFE3.
Optogenetic tools such as a BphP1–PpsR2 pair can be harnessed to exert spatiotemporal control over signaling pathways or transcriptional events. The BphP1–PpsR2 system is activated by near-infrared light, making it suitable for in vivo applications.
Growth inhibition metrics allow for robust measurement of drug efficacy independent of variables such as cell growth rate, seeding density, and growth medium; they are a practical alternative to metrics such as IC50 and offer enhanced reproducibility.
Treating glycoproteins with household bleach releases N- and O-glycans for further structural probing. Bleach treatment of glycosphingolipids releases glycan nitriles.
A method and accompanying software tool enables automated modeling of large macromolecular complexes using experimental crosslinking mass spectrometry data as distance restraints, as demonstrated for the 17-subunit yeast RNA polymerase III complex.
Microfluidic reprogramming of human somatic cells to induced pluripotent stem cells is rapid and highly efficient, enabling large-scale derivation from patient cells as well as seamless on-chip differentiation without cell expansion.