Volume 17 Issue 5, May 2022

This protocol describes a streamlined cross-linking and immunoprecipitation sequencing approach for transcriptome-wide detection of RNA-binding protein localization that is complemented by a comprehensive bioinformatics pipeline.

Skin organoids are generated from human pluripotent stem cells in a three-dimensional in vitro culture system. The mature organoids contain stratified skin layers, pigmented hair follicles, sebaceous glands, Merkel cells and sensory neurons.

RASER-FISH is a DNA fluorescence in situ hybridization technique that uses exonuclease digestion instead of heat denaturation to generate single-stranded target DNA for efficient probe binding, enabling chromatin preservation for super-resolution microscopy.

This chromosome-engineering protocol generates heritable chromosomal rearrangements in A. thaliana; by combining SaCas9 with an egg-cell-specific promoter to facilitate heritable mutations, chromosomal rearrangements can be made and homozygous lines can be established.

Functional islet organoids that can be cultured long term are generated from coculture of endothelial cells and pancreatic islet progenitors isolated via flow cytometry from adult mice.