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

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...

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Autores principales: Angelopoulou, Andriani, Papaspyropoulos, Angelos, Papantonis, Argyris, Gorgoulis, Vassilis G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8938320/
https://www.ncbi.nlm.nih.gov/pubmed/35330963
http://dx.doi.org/10.1016/j.xpro.2022.101257
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author Angelopoulou, Andriani
Papaspyropoulos, Angelos
Papantonis, Argyris
Gorgoulis, Vassilis G.
author_facet Angelopoulou, Andriani
Papaspyropoulos, Angelos
Papantonis, Argyris
Gorgoulis, Vassilis G.
author_sort Angelopoulou, Andriani
collection PubMed
description 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).
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spelling pubmed-89383202022-03-23 CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells Angelopoulou, Andriani Papaspyropoulos, Angelos Papantonis, Argyris Gorgoulis, Vassilis G. STAR Protoc Protocol 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). Elsevier 2022-03-18 /pmc/articles/PMC8938320/ /pubmed/35330963 http://dx.doi.org/10.1016/j.xpro.2022.101257 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Angelopoulou, Andriani
Papaspyropoulos, Angelos
Papantonis, Argyris
Gorgoulis, Vassilis G.
CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
title CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
title_full CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
title_fullStr CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
title_full_unstemmed CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
title_short CRISPR-Cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
title_sort crispr-cas9-mediated induction of large chromosomal inversions in human bronchial epithelial cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8938320/
https://www.ncbi.nlm.nih.gov/pubmed/35330963
http://dx.doi.org/10.1016/j.xpro.2022.101257
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