Construction of YAC-based mammalian artificial chromosomes

• 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 Nature Publishing Group US 01.05.1998; Nature
• Subjects: Agriculture; Animals; Autoantigens; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; 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; Life Sciences; Methods. Procedures. Technologies; Polymerase Chain Reaction; research-article; 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
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt0598-431

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.