Microdissection-derived Murine Mcb Probes from Somatic Cell Hybrids

Trifonov, Vladimir A.; Karst, Constanze; Claussen, Uwe; Mrasek, Kristin; Michel, Susanne; Avner, Philip; Liehr, Thomas

doi:10.1369/jhc.4b6598.2005

Cited by 9 publications

(13 citation statements)

References 8 publications

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“…4). The chromosomal backbone was visualized using multicolor banding (mcb) fragment 4, from a library of eight microdissected overlapping fragments from the mouse chromosome 6, as described previously (22,23). Multicolor banding fragment 4 extends from domains C1 to C3 of mouse chromosome 6 and harbors the CD8α gene (Fig.…”

Section: Resultsmentioning

confidence: 99%

Replication allows inactivation of a knocked-in locus control region in inappropriate cell lineages

Menzel

Ktistaki

Tolaini

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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To study the influence of a locus control region (LCR) on the expression of a highly characterized, developmentally regulated locus, we have targeted the hCD2-LCR as a single copy into the endogenous mouse CD8 gene complex. Two knock-in mouse lines that differ in the integration site of the hCD2-LCR within the mCD8 gene complex were generated, and the influence on expression of the CD8 coreceptor was assessed. In these mice the normal developmental silencing of the CD8 genes in the CD4 lineage is deregulated, and the mice develop CD4 + cells that also express the CD8 genes. This is accompanied by the physical maintenance of the CD8 genes within an extended loop away from their subchromosomal territory. Further analysis of these mice revealed unexpected fluid chromatin dynamics, whereby the LCR can be initially dominant over the endogenous CD8 gene-repressive regulatory processes present in CD4 + cells but is continuously contested by them, resulting in the eventual inactivation of the inserted LCR, probably as a result of multiple rounds of replication.CD8 locus | hCD2 locus control region | knock-in | lymphocyte development | chromatin structure

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Section: Resultsmentioning

confidence: 99%

Replication allows inactivation of a knocked-in locus control region in inappropriate cell lineages

Menzel

Ktistaki

Tolaini

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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“…A murine multicolor banding probe set for chromosome 6 was generated as reported in (59). Originally, eight pcp probes were generated by glass needlebased chromosome microdissection (Fig.…”

Section: Generation Of the Mcb Library For Chromosomementioning

confidence: 99%

CD8 Locus Nuclear Dynamics during Thymocyte Development

Ktistaki

Garefalaki

Williams

et al. 2010

The Journal of Immunology

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Nuclear architecture and chromatin reorganization have recently been shown to orchestrate gene expression and act as key players in developmental pathways. To investigate how regulatory elements in the mouse CD8 gene locus are arranged in space and in relation to each other, three-dimensional fluorescence in situ hybridization and chromosome conformation capture techniques were employed to monitor the repositioning of the locus in relation to its subchromosomal territory and to identify long-range interactions between the different elements during development. Our data demonstrate that CD8 gene expression in murine lymphocytes is accompanied by the relocation of the locus outside its subchromosomal territory. Similar observations in the CD4 locus point to a rather general phenomenon during T cell development. Furthermore, we show that this relocation of the CD8 gene locus is associated with a clustering of regulatory elements forming a tight active chromatin hub in CD8-expressing cells. In contrast, in nonexpressing cells, the gene remains close to the main body of its chromosomal domain and the regulatory elements appear not to interact with each other.

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“…For cross-species hybridizations, mcb probe sets for chromosomes 3, 6, 18, and 19 available at the Institute of Human Genetics and Anthropology (Jena), were applied as previously described Trifonov et al 2005, Karst et al 2006. A twofold-increased probe concentration and hybridization for up to 96 h was necessary for some hybridization.…”

Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

“…Recently, we described the generation of similar libraries for murine multicolour banding (mcb-the lower case is used here to distinguish these probes from MCB with human probes; Trifonov et al 2005;Karst et al 2006), as also reported by Benedek et al (2004). Here, we applied four selected autosomal mcb probes to study the evolution of chromosomes 3, 6, 18, and 19 in nine muroid species, belonging to subfamilies Murinae (Norway rat, striped field mouse), Cricetinae (golden, Chinese, greater long-tailed hamsters), and Arvicolinae (tundra vole, southern vole, collared lemming, mole vole).…”

Section: Introductionmentioning

confidence: 99%

New insights into the karyotypic evolution in muroid rodents revealed by multicolor banding applying murine probes

et al. 2010

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Muroid rodents are composed of a wide range of species characterized by extensive karyotypic evolution. Even if this group includes such important laboratory animal models as domestic mouse (Mus musculus), Norway rat (Rattus norvegicus), Chinese hamster (Cricetulus griseus), and golden hamster (Mesocricetus auratus), comparative cytogenetic studies between rodents are difficult due to the characteristic rapid karyotypic evolution. Molecular cytogenetic methods can help resolve problems of comparing muroid chromosomes. Here, we used cross-species comparative multicolour banding with probes obtained from mouse chromosomes 3, 6, 18, and 19 to study the karyotypes of nine muroid species from the three subfamilies Murinae, Cricetinae, and Arvicolinae. Results from multicolour banding with these murine probes (mcb) allowed us to improve the comparative homology maps between these species and to obtain new insights into their karyotypic evolution. We identified evolutionary conserved chromosomal breakpoints and revealed four previously unrecognized homologous segments, four inversions, and 14 evolutionary new centromeres in the nine muroid species studied. We found Mus apomorphic rearrangements, not seen in other muroids, and defined several subfamily specific chromosome breaks, characteristic for Arvicolinae and Cricetinae. We show that mcb libraries are an effective tool both for the cytogenetic characterisation of important laboratory models such as the rat and hamster as well as elucidating the complex phylogenomics relationships of muroids.

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Microdissection-derived Murine Mcb Probes from Somatic Cell Hybrids

Cited by 9 publications

References 8 publications

Replication allows inactivation of a knocked-in locus control region in inappropriate cell lineages

Replication allows inactivation of a knocked-in locus control region in inappropriate cell lineages

CD8 Locus Nuclear Dynamics during Thymocyte Development

New insights into the karyotypic evolution in muroid rodents revealed by multicolor banding applying murine probes

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