Volume 18 Issue 7, July 2023

The authors present a protocol for genome-wide clustered regularly interspaced short palindromic repeat screening of three-dimensional neurospheres.

We present a protocol for the encapsulation and stabilization of enzymes by using hydrogen-bonded organic frameworks (HOFs)—enzyme@HOF-101—along with procedures for their material characterization and testing of their biocatalytic performance.

A step-by-step guide providing biomedical researchers instructions for implementing spatial- and Fourier-domain ptychography to existing setups with minimal hardware modifications.

Cellulose is a burgeoning feedstock for more sustainable alternatives to fossil fuel-based materials. This protocol uses a cellulose-dissolving ionic liquid electrolyte with nuclear magnetic resonance methods to provide high-resolution structural analysis of cellulose.

The authors describe spiral volumetric optoacoustic tomography for the visualization of contrast agent perfusion and biodistribution in mice, featuring excellent scalability to achieve spatial resolution down to 90 µm or whole-body scans in <2 s.

A protocol for the generation of medulloblastoma and high-grade glioma organoids by electroporating, respectively, cerebellum and forebrain organoids, along with procedures for their use in drug screening, lineage tracing, co-culture and in vivo assays.

This protocol enables ex vivo immunocapture of cell-type-specific mitochondria directly from their tissue context in heterogeneous tissues such as the nervous system, using MitoTag reporter mice.

Detect-seq (dU-detection enabled by C-to-T transition during sequencing) enables off-target evaluation of programmable cytosine base editors. DNA is chemically labeled, and the editing intermediate dU is enriched for sequencing by biotin pull-down.

The authors present a FACS-based protocol for the purification of human skeletal stem cell lineages from a variety of tissue sources, alongside in vitro and in vivo skeletogenic functional assays, human xenograft mouse models and single-cell RNA sequencing analysis.

A protocol for the directed differentiation of human pluripotent stem cells into distal lung organoids that can be used to model interstitial lung disease, viral infection and human lung development, along with procedures for their analysis.

We present the design, assembly and implementation of a molecular-electromechanical system composed of a tetrahedral DNA nanostructure immobilized on a graphene field-effect transistor for unamplified detection of trace amounts of analytes in biofluids.

ProTracer is a genetic lineage–tracing technology that uses two recombinases to allow lifelong, noninvasive monitoring of cell proliferation for specific cell lineages in live animals.

Here the authors describe a protocol for generating bacterial cell lysates and freeze-dried, cell-free reactions for on-demand synthesis of glycoprotein therapeutics and vaccines, as well as aglycosylated proteins (for example, conjugate vaccine carrier proteins).