Aneuploidy, the presence of chromosome gains or losses, is a hallmark of cancer. Here, we describe KaryoCreate (karyotype CRISPR-engineered aneuploidy technology), a system that enables the generation of chromosome-specific aneuploidies by co-expression of an sgRNA targeting chromosome-specific CENPA-binding ɑ-satellite repeats together with dCas9 fused to mutant KNL1. We design unique and highly specific sgRNAs for 19 of the 24 chromosomes. Expression of these constructs leads to missegregation and induction of gains or losses of the targeted chromosome in cellular progeny, with an average efficiency of 8% for gains and 12% for losses (up to 20%) validated across 10 chromosomes. Using KaryoCreate in colon epithelial cells, we show that chromosome 18q loss, frequent in gastrointestinal cancers, promotes resistance to TGF-β, likely due to synergistic hemizygous deletion of multiple genes. Altogether, we describe an innovative technology to create and study chromosome missegregation and aneuploidy in the context of cancer and beyond.

KaryoCreate: A CRISPR-based technology to study chromosome-specific aneuploidy by targeting human centromeres / N. Bosco, A. Goldberg, X. Zhao, J.C. Mays, P. Cheng, A.F. Johnson, J.J. Bianchi, C. Toscani, E. Di Tommaso, L. Katsnelson, D. Annuar, S. Mei, R.E. Faitelson, I.Y. Pesselev, K.S. Mohamed, A. Mermerian, E.M. Camacho-Hernandez, C.A. Gionco, J. Manikas, Y. Tseng, Z. Sun, S. Fani, S. Keegan, S.M. Lippman, D. Fenyö, S. Giunta, S. Santaguida, T. Davoli. - In: CELL. - ISSN 1097-4172. - 186:9(2023 Apr 27), pp. 1985-2001. [10.1016/j.cell.2023.03.029]

KaryoCreate: A CRISPR-based technology to study chromosome-specific aneuploidy by targeting human centromeres

C. Toscani;S. Santaguida
Penultimo
;
2023

Abstract

Aneuploidy, the presence of chromosome gains or losses, is a hallmark of cancer. Here, we describe KaryoCreate (karyotype CRISPR-engineered aneuploidy technology), a system that enables the generation of chromosome-specific aneuploidies by co-expression of an sgRNA targeting chromosome-specific CENPA-binding ɑ-satellite repeats together with dCas9 fused to mutant KNL1. We design unique and highly specific sgRNAs for 19 of the 24 chromosomes. Expression of these constructs leads to missegregation and induction of gains or losses of the targeted chromosome in cellular progeny, with an average efficiency of 8% for gains and 12% for losses (up to 20%) validated across 10 chromosomes. Using KaryoCreate in colon epithelial cells, we show that chromosome 18q loss, frequent in gastrointestinal cancers, promotes resistance to TGF-β, likely due to synergistic hemizygous deletion of multiple genes. Altogether, we describe an innovative technology to create and study chromosome missegregation and aneuploidy in the context of cancer and beyond.
English
CRISPR; DNA damage; aneuploidy; cancer; centromere; chromosome gains and losses; chromosome missegregation; kinetochore; mitosis
Settore BIO/11 - Biologia Molecolare
Articolo
Esperti anonimi
Pubblicazione scientifica
Goal 3: Good health and well-being
27-apr-2023
Elsevier
186
9
1985
2001
17
Pubblicato
Periodico con rilevanza internazionale
pubmed
scopus
crossref
NON aderisco
info:eu-repo/semantics/article
KaryoCreate: A CRISPR-based technology to study chromosome-specific aneuploidy by targeting human centromeres / N. Bosco, A. Goldberg, X. Zhao, J.C. Mays, P. Cheng, A.F. Johnson, J.J. Bianchi, C. Toscani, E. Di Tommaso, L. Katsnelson, D. Annuar, S. Mei, R.E. Faitelson, I.Y. Pesselev, K.S. Mohamed, A. Mermerian, E.M. Camacho-Hernandez, C.A. Gionco, J. Manikas, Y. Tseng, Z. Sun, S. Fani, S. Keegan, S.M. Lippman, D. Fenyö, S. Giunta, S. Santaguida, T. Davoli. - In: CELL. - ISSN 1097-4172. - 186:9(2023 Apr 27), pp. 1985-2001. [10.1016/j.cell.2023.03.029]
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N. Bosco, A. Goldberg, X. Zhao, J.C. Mays, P. Cheng, A.F. Johnson, J.J. Bianchi, C. Toscani, E. Di Tommaso, L. Katsnelson, D. Annuar, S. Mei, R.E. Fai...espandi
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/1018028
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