Protocol for lentiviral engineering and multi-omic characterization of human kidney tubuloids

Organoids derived from normal human tissue have proven to be effective preclinical models for studying tumor progression through the introduction of driver mutations. In this protocol, we outline techniques for disease modeling using organoids derived from healthy kidney tissue, also known as tubuloids. We provide step-by-step instructions for the establishment and passaging of tubuloids, lentiviral transduction, processing for histological analysis, and preparation for CUT&RUN and single-cell transcriptomic profiling. For complete details on the use and execution of this protocol, please refer to Ganpat et al.¹.