The chromosomes of CHO, an aneuploid Chinese hamster cell line: G-band, C-band, and autoradiographic analyses

Deaven, Larry L.; Petersen, D.F.

doi:10.1007/bf00319690

Cited by 241 publications

(133 citation statements)

References 27 publications

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“…4B), where the painted karyotype clearly shows distinctive chromosomal aberrations, including both translocations and numerical changes. These changes are concurrent with previously discovered aberrations using G‐Banding and BAC hybridization reports 14, 15.…”

Section: Resultssupporting

confidence: 85%

ChromaWizard: An open source image analysis software for multicolor fluorescence in situ hybridization analysis

et al. 2018

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Multicolor image analysis finds its applications in a broad range of biological studies. Specifically, multiplex fluorescence in situ hybridization (M‐FISH) for chromosome painting facilitates the analysis of individual chromosomes in complex metaphase spreads and is widely used to detect both numerical and structural aberrations. While this is well established for human and mouse karyotypes, for which species sophisticated software and analysis tools are available, other organisms and species are less well served. Commercially available software is proprietary and not easily adaptable to other karyotypes. Therefore, a publically available open source software that combines flexibility and customizable functionalities is needed. Here we present such a tool called “ChromaWizard” which is based on popular scientific image analysis libraries (OpenCV, scikit‐image, and NumPy). We demonstrate its functionality on the example of primary Chinese hamster (Cricetulus griseus) fibroblasts metaphase spreads and on Chinese Hamster Ovary cell lines known for the large number of chromosomal rearrangements. The application can be easily adapted to any kind of available labeling kits and is https://dict.leo.org/englisch-deutsch/independent of the used organism and instrumentation. It allows direct inspection of the original hybridization signals and enables either manual or automatic assignment of colors, making it a functional and versatile tool that can be used also for other multicolor applications.

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Section: Resultssupporting

confidence: 85%

ChromaWizard: An open source image analysis software for multicolor fluorescence in situ hybridization analysis

et al. 2018

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“…Numerous studies have shown large chromosomal rearrangements in CHO cells, using banding techniques [41][42][43] and fluorescence in situ hybridization 8,[44][45][46][47][48] . These approaches identified large translocations in CHO cells, providing a coarse-grained view of genomic variations in these unstable genomes.…”

Section: Discussionmentioning

confidence: 99%

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome

Lewis¹,

et al. 2013

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Chinese hamster ovary (CHO) cells, first isolated in 1957, are the preferred production host for many therapeutic proteins. Although genetic heterogeneity among CHO cell lines has been well documented, a systematic, nucleotide-resolution characterization of their genotypic differences has been stymied by the lack of a unifying genomic resource for CHO cells. Here we report a 2.4-Gb draft genome sequence of a female Chinese hamster, Cricetulus griseus, harboring 24,044 genes. We also resequenced and analyzed the genomes of six CHO cell lines from the CHO-K1, DG44 and CHO-S lineages. This analysis identified hamster genes missing in different CHO cell lines, and detected >3.7 million single-nucleotide polymorphisms (SNPs), 551,240 indels and 7,063 copy number variations. Many mutations are located in genes with functions relevant to bioprocessing, such as apoptosis. The details of this genetic diversity highlight the value of the hamster genome as the reference upon which CHO cells can be studied and engineered for protein production.

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“…Moreover, it has been maintained in a CHO cell line that has been in culture since 1957 (Puck et al, 1958). During adaptation to tissue culture conditions, many chromosomal changes took place in this cell line resulting in its current karyotype of 21-22 nonidentical chromosomes (Deaven and Petersen, 1973). In this process, many genes were lost or became haploid (Siminovitch, 1976).…”

Section: Discussionmentioning

confidence: 99%

A Ubiquitous β-tubulin Disrupts Microtubule Assembly and Inhibits Cell Proliferation

Bhattacharya

Cabral

2004

MBoC

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Vertebrate tubulin is encoded by a multigene family that produces distinct gene products, or isotypes, of both the ␣-and ␤-tubulin subunits. The isotype sequences are conserved across species supporting the hypothesis that different isotypes subserve different functions. To date, however, most studies have demonstrated that tubulin isotypes are freely interchangeable and coassemble into all classes of microtubules. We now report that, in contrast to other isotypes, overexpression of a mouse class V ␤-tubulin cDNA in mammalian cells produces a strong, dose-dependent disruption of microtubule organization, increased microtubule fragmentation, and a concomitant reduction in cellular microtubule polymer levels. These changes also disrupt mitotic spindle assembly and block cell proliferation. Consistent with diminished microtubule assembly, there is an increased tolerance for the microtubule stabilizing drug, paclitaxel, which is able to reverse many of the effects of class V ␤-tubulin overexpression. Moreover, transfected cells selected in paclitaxel exhibit increased expression of class V ␤-tubulin, indicating that this isotype is responsible for the drug resistance. The results show that class V ␤-tubulin is functionally distinct from other tubulin isotypes and imparts unique properties on the microtubules into which it incorporates. INTRODUCTIONMicrotubules are essential filamentous structures in eukaryotic cells where they are responsible for the directed movement of vesicles, the organization of the endoplasmic reticulum and Golgi apparatus in the cytoplasm, and the equipartitioning of chromosomes before cell division. The organelles are assembled from heterodimers of ␣-and ␤-tubulin, which polymerize in a head-to-tail manner to form linear protofilaments, and these associate laterally into tubular structures that normally consist of 13 protofilaments. Vertebrate ␣-and ␤-tubulins are each encoded by a 6-to 7-member multigene family that produces highly homologous and conserved gene products that differ most radically in their last 10 -15 amino acids (Sullivan, 1988;Luduena, 1998). These carboxyl-terminal sequences have been used to assign ␤-tubulin gene products to seven distinct classes (Lopata and Cleveland, 1987). Each of these classes (hereafter referred to as ␤1, ␤2, ␤3, ␤4a, ␤4b, ␤5, and ␤6) defines a ␤-tubulin isotype that differs significantly from other isotypes within the same organism, but differs very little from the same isotype in other vertebrate species.With the discovery that tubulin proteins in an organism are heterogeneous, a hypothesis was formulated suggesting that different tubulin proteins might perform different functions in the cell (Fulton and Simpson, 1976). In the intervening years, however, most of the experimental and genetic evidence has argued that different ␤-tubulins coassemble freely into all cellular microtubules (see Joshi and Cleveland, 1990; Luduena, 1998 for reviews). In gene replacement experiments using the fungal organism, Aspergillus nidulans, for example, May demonst...

show abstract

The chromosomes of CHO, an aneuploid Chinese hamster cell line: G-band, C-band, and autoradiographic analyses

Cited by 241 publications

References 27 publications

ChromaWizard: An open source image analysis software for multicolor fluorescence in situ hybridization analysis

ChromaWizard: An open source image analysis software for multicolor fluorescence in situ hybridization analysis

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome

A Ubiquitous β-tubulin Disrupts Microtubule Assembly and Inhibits Cell Proliferation

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