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

## TL;DR

This paper provides a detailed protocol for creating and analyzing kidney tubuloids using lentiviral engineering and multi-omic techniques.

## Contribution

The paper introduces a comprehensive protocol for disease modeling using kidney tubuloids with multi-omic profiling.

## Key findings

- A protocol is outlined for establishing and passaging tubuloids derived from healthy kidney tissue.
- Lentiviral transduction methods are described for introducing genetic modifications into tubuloids.
- The protocol includes steps for CUT&RUN and single-cell transcriptomic profiling of tubuloids.

## Abstract

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

•Protocol for establishment, passaging, and lentiviral transduction of tubuloids•Steps for histological and immunohistochemical staining•Preparation for CUT&RUN and single-cell transcriptomic profiling•Applicable to organoids derived from healthy and tumor tissues

Protocol for establishment, passaging, and lentiviral transduction of tubuloids

Steps for histological and immunohistochemical staining

Preparation for CUT&RUN and single-cell transcriptomic profiling

Applicable to organoids derived from healthy and tumor tissues

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Organoids derived from normal human tissues 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.