Protocol for generation and engineering of thyroid cell lineages using CRISPR-Cas9 editing to recapitulate thyroid cancer histotype progression

Vincenzo Davide Pantina; Francesco Verona; Alice Turdo; Veronica Veschi; Chiara Modica; Melania Lo Iacono; Miriam Gaggianesi; Sebastiano Di Bella; Matilde Todaro; Simone Di Franco; Giorgio Stassi

doi:10.1016/j.xpro.2024.103263

Protocol for generation and engineering of thyroid cell lineages using CRISPR-Cas9 editing to recapitulate thyroid cancer histotype progression

STAR Protoc. 2024 Sep 20;5(3):103263. doi: 10.1016/j.xpro.2024.103263. Epub 2024 Aug 10.

Affiliations

¹ Department of Precision Medicine in Medical, Surgical and Critical Care, University of Palermo, 90100 Palermo, Italy.
² Department of Health Promotion, Mother and Child Care, Internal Medicine and Medical Specialties, University of Palermo, 90100 Palermo, Italy.
³ Department of Precision Medicine in Medical, Surgical and Critical Care, University of Palermo, 90100 Palermo, Italy; Department of Molecular Medicine, University La Sapienza, 00161 Rome, Italy.
⁴ Department of Health Promotion, Mother and Child Care, Internal Medicine and Medical Specialties, University of Palermo, 90100 Palermo, Italy; A.O.U.P. "Paolo Giaccone", University of Palermo, 90100 Palermo, Italy.
⁵ Department of Precision Medicine in Medical, Surgical and Critical Care, University of Palermo, 90100 Palermo, Italy. Electronic address: [email protected].
⁶ Department of Precision Medicine in Medical, Surgical and Critical Care, University of Palermo, 90100 Palermo, Italy. Electronic address: [email protected].

Abstract

Thyroid carcinoma represents the first malignancy among the endocrine organs. Investigating the cellular hierarchy and the mechanisms underlying the initiation of thyroid carcinoma is crucial in thyroid cancer research. Here, we present a protocol for deriving thyroid cell lineage from human embryonic stem cells. We also describe steps for engineering thyroid progenitor cells utilizing CRISPR-Cas9 technology, which can be used to perform in vivo studies, thus facilitating the development of representative thyroid tumorigenesis models. For complete details on the use and execution of this protocol, please refer to Veschi et al.¹.

Keywords: CRISPR; Cancer; Cell Differentiation; Cell culture.

MeSH terms

CRISPR-Cas Systems* / genetics
Cell Lineage* / genetics
Disease Progression
Gene Editing* / methods
Human Embryonic Stem Cells / cytology
Human Embryonic Stem Cells / metabolism
Humans
Thyroid Gland* / cytology
Thyroid Gland* / metabolism
Thyroid Gland* / pathology
Thyroid Neoplasms* / genetics
Thyroid Neoplasms* / pathology