KaryoCreate: A CRISPR-based technology to study chromosome-specific aneuploidy by targeting human centromeres.
CRISPR
DNA damage
aneuploidy
cancer
centromere
chromosome gains and losses
chromosome missegregation
kinetochore
mitosis
Journal
Cell
ISSN: 1097-4172
Titre abrégé: Cell
Pays: United States
ID NLM: 0413066
Informations de publication
Date de publication:
27 04 2023
27 04 2023
Historique:
received:
16
06
2022
revised:
17
11
2022
accepted:
23
03
2023
pmc-release:
27
04
2024
medline:
1
5
2023
pubmed:
20
4
2023
entrez:
19
04
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 37075754
pii: S0092-8674(23)00326-4
doi: 10.1016/j.cell.2023.03.029
pmc: PMC10676289
mid: NIHMS1893040
pii:
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1985-2001.e19Subventions
Organisme : NCI NIH HHS
ID : P30 CA016087
Pays : United States
Organisme : NCI NIH HHS
ID : P50 CA097007
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM136542
Pays : United States
Organisme : NCI NIH HHS
ID : P01 CA106451
Pays : United States
Organisme : NIDCR NIH HHS
ID : R01 DE026644
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA023100
Pays : United States
Organisme : Cancer Research UK
ID : C5470/A27144
Pays : United Kingdom
Organisme : NCI NIH HHS
ID : R37 CA248631
Pays : United States
Organisme : NCI NIH HHS
ID : R00 CA212621
Pays : United States
Organisme : NCI NIH HHS
ID : U24 CA210972
Pays : United States
Commentaires et corrections
Type : CommentIn
Informations de copyright
Copyright © 2023 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests T.D. is on the Scientific Advisory Board (SAB) of io9. S.M.L. is a co-founder of io9 and a member of Biological Dynamics, Inc. SAB. T.D. and N.B. are inventors on patent application no. 63/375,181.
Références
Cell. 2015 Dec 17;163(7):1641-54
pubmed: 26687355
Bioinformatics. 2009 Jul 15;25(14):1754-60
pubmed: 19451168
Bioinformatics. 2014 Apr 1;30(7):923-30
pubmed: 24227677
Cold Spring Harb Perspect Biol. 2014 Jul 01;6(7):a015826
pubmed: 24984773
Nat Commun. 2018 Oct 18;9(1):4340
pubmed: 30337534
Genome Res. 2017 May;27(5):849-864
pubmed: 28396521
Nat Genet. 1996 Jul;13(3):343-6
pubmed: 8673134
Science. 2013 Feb 15;339(6121):819-23
pubmed: 23287718
Science. 2022 Apr;376(6588):44-53
pubmed: 35357919
Cell. 2013 Jul 3;154(1):47-60
pubmed: 23827674
Cancer Cell. 2018 Apr 9;33(4):676-689.e3
pubmed: 29622463
Biology (Basel). 2017 Jan 24;6(1):
pubmed: 28125021
Genome Biol. 2015 Feb 27;16:49
pubmed: 25887352
Nat Biotechnol. 2016 Feb;34(2):184-191
pubmed: 26780180
EMBO J. 2020 Jan 15;39(2):e102924
pubmed: 31750958
Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70
pubmed: 21307310
Nat Genet. 2019 Apr;51(4):705-715
pubmed: 30833795
Cell. 2013 Dec 19;155(7):1479-91
pubmed: 24360272
Bioinformatics. 2014 Aug 1;30(15):2114-20
pubmed: 24695404
Genome Biol. 2014;15(12):550
pubmed: 25516281
Structure. 2018 Oct 2;26(10):1327-1336.e4
pubmed: 30100357
Elife. 2022 Sep 21;11:
pubmed: 36129397
Trends Cell Biol. 2011 Jun;21(6):374-81
pubmed: 21306900
Dis Colon Rectum. 2004 Sep;47(9):1467-82
pubmed: 15486743
PeerJ. 2014 Jun 19;2:e453
pubmed: 25024921
Cell. 2013 Nov 7;155(4):948-62
pubmed: 24183448
Annu Rev Genomics Hum Genet. 2006;7:301-13
pubmed: 16756479
Cell. 2021 Jun 24;184(13):3573-3587.e29
pubmed: 34062119
Sci Rep. 2019 Sep 16;9(1):13292
pubmed: 31527759
Proc Natl Acad Sci U S A. 2012 Feb 7;109(6):1979-84
pubmed: 22308327
Nat Methods. 2012 Jun 28;9(7):676-82
pubmed: 22743772
Cell. 1996 Aug 23;86(4):543-52
pubmed: 8752209
J Cell Biol. 2010 Mar 22;188(6):809-20
pubmed: 20231380
Mol Syst Biol. 2012;8:608
pubmed: 22968442
Science. 2014 Jun 20;344(6190):1396-401
pubmed: 24925914
Curr Biol. 2001 Sep 18;11(18):1462-7
pubmed: 11566107
Nat Biotechnol. 2014 Dec;32(12):1262-7
pubmed: 25184501
Cell. 2015 May 7;161(4):933-45
pubmed: 25957691
Chromosome Res. 2012 Jul;20(5):621-33
pubmed: 22801774
Science. 2022 Apr;376(6588):eabl4178
pubmed: 35357911
Elife. 2020 Nov 11;9:
pubmed: 33174837
Cell. 2006 Sep 8;126(5):995-1004
pubmed: 16959577
Bioinformatics. 2016 Sep 15;32(18):2847-9
pubmed: 27207943
Nature. 2010 Feb 18;463(7283):899-905
pubmed: 20164920
Dev Cell. 2017 Aug 7;42(3):226-240.e6
pubmed: 28787590
Br J Cancer. 2006 Dec 4;95(11):1562-7
pubmed: 17088901
J Cell Biol. 2010 Jul 12;190(1):25-34
pubmed: 20624899
Cell Rep. 2021 Nov 9;37(6):109818
pubmed: 34758321
Proc Natl Acad Sci U S A. 2012 May 22;109(21):8212-7
pubmed: 22566646
Science. 2018 Jan 5;359(6371):108-114
pubmed: 29170278
Nature. 2015 May 7;521(7550):43-7
pubmed: 25924068
Genes Dev. 2021 Aug 1;35(15-16):1079-1092
pubmed: 34266888
Proc Natl Acad Sci U S A. 2005 Oct 25;102(43):15545-50
pubmed: 16199517
Nat Methods. 2014 Aug;11(8):783-784
pubmed: 25075903
Cell. 2018 Apr 5;173(2):400-416.e11
pubmed: 29625055
Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9719-23
pubmed: 11481457
J Cell Biol. 2001 Apr 30;153(3):517-27
pubmed: 11331303
Data Brief. 2019 Mar 08;24:103708
pubmed: 30989093
Bioinformatics. 2013 Jan 1;29(1):15-21
pubmed: 23104886
CRISPR J. 2019 Dec;2(6):406-416
pubmed: 31742432
Neoplasia. 2011 Apr;13(4):348-57
pubmed: 21472139
Nat Cell Biol. 2017 Jan;19(1):68-75
pubmed: 27918550
Proc Natl Acad Sci U S A. 1981 Oct;78(10):6349-53
pubmed: 6947234
J Cell Biol. 2010 Jul 12;190(1):73-87
pubmed: 20624901
Nat Biotechnol. 2021 May;39(5):599-608
pubmed: 33462507
EMBO J. 2023 May 15;42(10):e111559
pubmed: 37038978
Proc Natl Acad Sci U S A. 2021 Mar 9;118(10):
pubmed: 33653953
Exp Cell Res. 2020 Apr 15;389(2):111896
pubmed: 32035947
Nature. 2012 Jul 18;487(7407):330-7
pubmed: 22810696
Science. 2014 Jan 3;343(6166):80-4
pubmed: 24336569
Nat Genet. 2017 Dec;49(12):1779-1784
pubmed: 29083409
Cell. 2006 Dec 1;127(5):969-82
pubmed: 17129782
Biochem Soc Trans. 2009 Oct;37(Pt 5):976-80
pubmed: 19754435
Nature. 2020 Nov;587(7832):126-132
pubmed: 32879494
Cell. 2014 Oct 23;159(3):647-61
pubmed: 25307932
Science. 2016 Apr 8;352(6282):189-96
pubmed: 27124452
Curr Biol. 2015 Oct 19;25(20):R1002-18
pubmed: 26485365
Proc Natl Acad Sci U S A. 2014 Sep 16;111(37):13409-14
pubmed: 25197050
Nat Protoc. 2013 Nov;8(11):2281-2308
pubmed: 24157548
Genome Biol. 2017 Nov 24;18(1):224
pubmed: 29178945
Curr Opin Genet Dev. 1991 Dec;1(4):509-14
pubmed: 1822282
Proc Natl Acad Sci U S A. 2021 May 11;118(19):
pubmed: 33952700
Cell. 2014 Mar 13;156(6):1298-1311
pubmed: 24630729
Cell. 2000 Oct 13;103(2):295-309
pubmed: 11057902