Mouse A9 cells containing single human chromosomes for analysis of genomic imprinting

To develop an systematic in vitro approach for the study of genomic imprinting, we generated a new library of human/mouse A9 monochromosomal hybrids. We used whole cell fusion and microcell-mediated chromosome transfer to generate A9 hybrids containing a single, intact, bsr-tagged human chromosome d...

Full description

Saved in:
Bibliographic Details
Published in:DNA research 1999, Vol.6 (3), p.165-172
Main Authors: Kugoh, H, Mitsuya, K, Meguro, M, Shigenami, K, Schulz, T C, Oshimura, M
Format: Article
Language:English
Subjects:
Animals
Cell Line
Chromosomes, Human - genetics
Female
Gene Expression
Genomic Imprinting - genetics
Humans
Hybrid Cells
In Situ Hybridization, Fluorescence
Karyotyping
Male
Mice
Microsatellite Repeats
Polymorphism, Genetic - genetics
Transfection
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:To develop an systematic in vitro approach for the study of genomic imprinting, we generated a new library of human/mouse A9 monochromosomal hybrids. We used whole cell fusion and microcell-mediated chromosome transfer to generate A9 hybrids containing a single, intact, bsr-tagged human chromosome derived from primary fibroblasts. A9 hybrids were identified that contained either human chromosome 1, 2, 4, 5, 7, 8, 10, 11, 15, 18, 20, or X. The parental origin of these chromosomes was determined by polymorphic analysis using microsatellite markers, and matched hybrids containing maternal and paternal chromosomes were identified for chromosomes 5, 10, 11 and 15. The imprinted gene KVLQT1 on human chromosome 11p15.5 was expressed exclusively from the maternal chromosome in A9 hybrids, and the parental-origin-specific expression patterns of several other imprinted genes were also maintained. This library of human monochromosomal hybrids is a valuable resource for the mapping and cloning of human genes and is a novel in vitro system for the screening of imprinted genes and for their functional analysis.
ISSN:1340-2838
DOI:10.1093/dnares/6.3.165