Construction of YAC-based mammalian artificial chromosomes

To construct a mammalian artificial chromosome (MAC), telomere repeats and selectable markers were introduced into a 100 kb yeast artificial chromosome (YAC) containing human centromeric DNA. This YAC, which has a regular repeat structure of alpha-satellite DNA and centromere protein B (CENP-B) boxe...

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Bibliographic Details
Published in:	Nature biotechnology Vol. 16; no. 5; pp. 431 - 439
Main Authors:	Ikeno, Masashi, Grimes, Brenda, Okazaki, Tuneko, Nakano, Megumi, Saitoh, Kaori, Hoshino, Harumi, McGill, Niolette I, Cooke, Howard, Masumoto, Hiroshi
Format:	Journal Article
Language:	English
Published:	New York, NY Nature 01-05-1998
Subjects:	Animals Autoantigens Biological and medical sciences Biotechnology Cell Line Cell Line, Transformed Centromere - genetics Centromere Protein B Chromosomal Proteins, Non-Histone - chemistry Chromosomal Proteins, Non-Histone - genetics Chromosomes - genetics Chromosomes, Artificial, Yeast - genetics Chromosomes, Human, Pair 21 - genetics Cloning, Molecular DNA, Satellite - chemistry DNA, Satellite - genetics DNA-Binding Proteins Fundamental and applied biological sciences. Psychology Genetic engineering Genetic technics Genetic Vectors - genetics Humans In Situ Hybridization, Fluorescence Kinetochores Methods. Procedures. Technologies Polymerase Chain Reaction Telomere - genetics Transfection Vectors (cloning, transfer, expression). Insertion sequences and transposons Human Alpha satellite DNA Vertebrata Cell line Mammalia Artificial chromosome Vector Structural analysis Genetic transfer
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Summary:	To construct a mammalian artificial chromosome (MAC), telomere repeats and selectable markers were introduced into a 100 kb yeast artificial chromosome (YAC) containing human centromeric DNA. This YAC, which has a regular repeat structure of alpha-satellite DNA and centromere protein B (CENP-B) boxes, efficiently formed MACs that segregated accurately and bound CENP-B, CENP-C, and CENP-E. The MACs appear to be about 1-5 Mb in size and contain YAC multimers. Structural analyses suggest that the MACs have not acquired host sequences and were formed by a de novo mechanism. The accurate segregation of the MACs suggests they have potential as vectors for introducing genes into mammals.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	1087-0156 1546-1696
DOI:	10.1038/nbt0598-431