CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells

Andriani Angelopoulou; Angelos Papaspyropoulos; Argyris Papantonis; Vassilis G. Gorgoulis

STAR Protocols (Jun 2022)

CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells

Andriani Angelopoulou,
Angelos Papaspyropoulos,
Argyris Papantonis,
Vassilis G. Gorgoulis

Affiliations

Andriani Angelopoulou: Molecular Carcinogenesis Group, Department of Histology and Embryology, Faculty of Medicine, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
Angelos Papaspyropoulos: Molecular Carcinogenesis Group, Department of Histology and Embryology, Faculty of Medicine, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
Argyris Papantonis: Translational Epigenetics Group, Institute of Pathology, University Medical Center Göttingen, 37075 Göttingen, Germany; Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany; Corresponding author
Vassilis G. Gorgoulis: Molecular Carcinogenesis Group, Department of Histology and Embryology, Faculty of Medicine, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Division of Cancer Sciences, School of Medical Sciences, Faculty of Biology, Medicine & Health, University of Manchester, M20 4GJ Manchester, UK; Center for New Biotechnologies and Precision Medicine, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Faculty of Health and Medical Sciences, University of Surrey, GU2 7YH Surrey, UK; Corresponding author

Journal volume & issue: Vol. 3, no. 2
p. 101257

Abstract

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Summary: The in vitro recapitulation of chromosomal rearrangements is a necessary tool for understanding malignancy at the molecular level. Here, we describe the targeted induction of a large chromosomal inversion (>3.7 Mbp) through CRISPR-Cas9-mediated genome editing. As inversions occur at low frequency following Cas9 cleavage, we provide a detailed screening approach of FACS-sorted, single-cell-derived clonal human bronchial epithelial cell (HBEC) cultures. The protocol provided is tailored to HBECs; however, it can be readily applied to additional adherent cellular models.For complete details on the use and execution of this protocol, please refer to Zampetidis et al. (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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