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The authors provide an expanded CRISPR–Cas9-assisted recombineering toolkit for rapid and efficient engineering of genetically intractable Pseudomonas aeruginosa isolates.
The authors describe a new base editing system—the transformer base editor—to induce efficient editing with no observable genome-wide or transcriptome-wide off-target mutations, both in mammalian cells and in mice.
Microbial pathogens develop resistance to clinically used drugs, and finding new therapeutics is an ongoing challenge. The photoimmuno-antimicrobial strategy described in this protocol is a general approach to target specific elimination.
Invasion–adhesion-directed expression sequencing adapts the 10x Genomics 5′ single-cell RNA sequencing protocol to enable generation of bacterial and eukaryotic DNA libraries to identify adherent or invasive bacteria and the associated host transcriptome at a single-cell level.
The authors provide a protocol to microscopically identify and isolate single maize meiocytes and pollen grains for RNA-seq with a modified version of CEL-Seq2. Staging of developmentally synchronized anthers is performed in parallel with expression analysis.
This protocol can be used to generate kidney organoids with elongated proximalized nephrons displaying improved proximal tubule maturity compared with those generated using standard kidney organoid protocols.
Procedures for the synthesis of physical double-network hydrogels to provide self-healing, pH-responsive, tissue-adhesive, antioxidant, photothermal and antibacterial properties. Used as wound dressings, the hydrogels can also be removed on demand.
This protocol provides structure-guided capsid library approaches to evolve new adeno-associated viral vectors for enhanced CNS gene delivery with altered tissue tropism, higher transduction efficiency and the ability to evade pre-existing humoral immunity.
This protocol describes how to prepare ordered mesoporous intermetallic nanomaterials with controlled compositions, morphologies/structures and phases via a general concurrent template strategy.
Purifying proteins by crystallization rather than chromatography is less expensive and more environmentally sustainable. This protocol describes how to determine the operational conditions for purifying monoclonal antibodies by template-assisted membrane crystallization.
This Protocol documents the extensive preparation required to adopt Neuropixels probes for use in patients for recording spiking activity from up to hundreds of cortical neurons and associating it with local field potentials.
Immobilization of enzymes in solid porous supports can enhance the recyclability and stability of enzymes. This protocol describes direct-immobilization methods to make enzyme-covalent organic framework biocomposites.
A correlative imaging approach in which matrix-assisted laser desorption ionization mass spectrometry imaging is followed by fluorescence in situ hybridization (metaFISH) for the spatial metabolomics identification of host–microbiome interactions, at single-cell resolution, via nucleic acid probes.
A simple and robust approach for the large-scale assembly of DNA-templated precise carbon nanotube semiconductor arrays with clean channel interfaces to build field-effect transistors for high-performance electronics.
This protocol for universal and proficient Pseudomonas recombineering uses phage-encoded homologous recombination–Cas3 systems, including SacB counterselection and Cre site-specific recombinase for two- or three-step seamless genome modification.
We present a protocol for generating mouse ESC-derived induced neuronal cells via lentiviral Tet-On inducible expression of NGN2 for the direct comparison of human and mouse neuronal populations.
This protocol describes the critical concepts, experimental standardization, operational principles and data analysis for using on-chip electrocatalytic microdevices.
This protocol is for using CapQuant, a mass spectrometry-based technique for accurate and sensitive quantification of the epitranscriptome of RNA caps using stable isotope-labeled cap nucleotides.
A protocol for setting up, calibrating and validating a semi-automated sample processing pipeline for cell-free RNA isolation from clinical samples using the Opentrons 1.0 or 2.0 open-source robotic platform.
A protocol for broad-spectrum, in vivo fluorescent labeling and tracking of extracellular matrix proteins through the systemic or local application of N-hydroxysuccinimide esters. This flexible approach can be adapted for a variety of organ systems and wounding models.