Inducing Specific Chromosome Mis-Segregation in Human Cells
Inducing Specific Chromosome Mis-Segregation in Human Cells
复制标题
诱导人体细胞中的特定染色体错误分离
DOI：
10.1101/2022.04.19.486691
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Tovini L
中科院分区：
文献类型：
--
作者：
Tovini L
癌细胞表现出持续的潜在染色体不稳定性，导致染色体错误分离、微核形成和染色体数目异常（非整倍性）。这些特征是几乎所有人类癌症所共有的，其中个别肿瘤类型有趣地表现出完整和亚染色体非整倍性的特征子集。迄今为止，很少有方法诱导特异性非整倍体存在，阻碍了调查功能的后果复发性非整倍体。此外，尽管存在一些具有特异性非整倍体的人类细胞系，但对特异性染色体不稳定事件的急性细胞反应仍然未知。因此，我们研究了使用核酸酶死亡的CRISPR-Cas9（dCas 9）作为特定基因组基因座的货物载体来破坏特定染色体的有丝分裂分离的可能性。我们招募着丝粒蛋白CENP-T的着丝粒成核结构域组装异位着丝粒附近的9号染色体，或染色体1的端粒。异位动粒组装导致靶染色体的染色体不稳定性和非整倍性增加，从而提供了创建“设计者核型”并在广泛的细胞类型中研究其直接下游细胞反应的潜力。总体而言，我们的发现为异位动粒生物学提供了新的见解，也代表了研究特定非整倍性和染色体错误分离事件在与非整倍性相关疾病中的作用的重要一步。
Cancer cells display persistent underlying chromosomal instability that results in chromosome mis-segregation, the formation of micronuclei, and abnormal numbers of chromosomes (aneuploidy). These features are common to nearly all human cancers, with individual tumour types intriguingly exhibiting characteristic subsets of whole, and sub-chromosomal aneuploidies. To date, few methods to induce specific aneuploidies at will exist, hampering the investigation of functional consequences of recurrent aneuploidies. Moreover, although some human cell lines with specific aneuploidies exist, theacutecellular responses to specific chromosomal instability events remain unknown. We therefore investigated the possibility of sabotaging the mitotic segregation of specific chromosomes using nuclease-dead CRISPR-Cas9 (dCas9) as a cargo carrier to specific genomic loci. We recruited the kinetochore-nucleating domain of centromere protein CENP-T to assemble ectopic kinetochores either near the centromere of chromosome 9, or the telomere of chromosome 1. Ectopic kinetochore assembly led to increased chromosome instability and aneuploidy of the target chromosomes, providing the potential to create ‘designer karyotypes’ and study their immediate downstream cellular responses in a wide range of cell types. Overall, our findings provide new insights into ectopic kinetochore biology, and also represent an important step towards investigating the role of specific aneuploidy and chromosome mis-segregation events in diseases associated with aneuploidy.
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影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
50.3
作者：
Taylor AM;Shih J;Ha G;Gao GF;Zhang X;Berger AC;Schumacher SE;Wang C;Hu H;Liu J;Lazar AJ;Cancer Genome Atlas Research Network;Cherniack AD;Beroukhim R;Meyerson M
通讯作者：
Meyerson M
影响因子：
12.3
作者：
van den Bos H;Spierings DC;Taudt AS;Bakker B;Porubský D;Falconer E;Novoa C;Halsema N;Kazemier HG;Hoekstra-Wakker K;Guryev V;den Dunnen WF;Foijer F;Tatché MC;Boddeke HW;Lansdorp PM
通讯作者：
Lansdorp PM
DOI：
10.1111/j.1749-6632.1975.tb19218.x
发表时间：
1975-01-01
影响因子：
5.2
作者：
BRINKLEY, BR;CARTWRIGHT, J
通讯作者：
CARTWRIGHT, J
影响因子：
16.6
作者：
Passerini V;Ozeri-Galai E;de Pagter MS;Donnelly N;Schmalbrock S;Kloosterman WP;Kerem B;Storchová Z
通讯作者：
Storchová Z