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

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...

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Bibliographic Details
Main Authors: Andriani Angelopoulou, Angelos Papaspyropoulos, Argyris Papantonis, Vassilis G. Gorgoulis
Format: Article
Language:English
Published: Elsevier 2022-06-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S266616672200137X
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Summary: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).
ISSN:2666-1667