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2002 Fiscal Year Final Research Report Summary

Creation of a mouse with human MHC by Chromosomal- and Developmental-engineering methods

Research Project

Project/Area Number 12554036
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 遺伝
Research InstitutionNational Institute of Genetics

Principal Investigator

IKEMURA Toshimichi  National Inst. of Genetics, Dept. of Popul. Genet., Professor, 集団遺伝研究系, 教授 (50025475)

Co-Investigator(Kenkyū-buntansha) FUKAGAWA Tatsuo  National Inst. of Genetics, Dept. of Popul. Genet, Associate Professor, 集団遺伝研究系, 助教授 (60321600)
Project Period (FY) 2000 – 2002
KeywordsArtificial Chromosomes / MHC / Chromosome Engineering / DT40 / Centromere / Developmental Engineering / Genome Engineering / Chromosome Vector
Research Abstract

Genetic and chromosomal engineering and transgenic animal technology have greatly contributed to progress of all fields of life sciences, including medicine, agriculture, and pharmacy. We developed chromosome engineering technique to manipulate a large domain of human chromosomes, by introducing several to several-tens Mb portions of human chromosome into chicken DT40 cells, in which frequencies of homologous recombination are several tens to hundreds times higher than those in mammals cell and therefore human chromosome can be efficiently manipulated. Centomere has important roles in production of artificial chromosome with functions properly regulated (Fukagawa et al., 2001 ; Nishihashi et al, 2002).
The human MHC locus (4 Mb) consists of three functional domains (classes I, II, and III). We found individual domains to replicate in different periods during S phase and transition of replication timing to occurs at boundaries of the functional domains. French group reported that in the … More case of the pig MHC locus, centromere exists in the boundary of the classes II and III. In this boundary in the human MHC locus we found characteristic sequences that have potentiality to form triple helix (Kanaya et al.,). We also found the triplex structure, which is predicted to modulate replication timing, was formed also within the nucleus and specially associated with centromere sequences (Ohno et al., 2002). Centromeric DNA that exists on the artificial chromosome was found in the late period during S phase, and the replication timing changed after integrating into a host's chromosome. To get artificial chromosomes with proper functions accurately regulated, it is important to clarify the centromeric structures and functions. We analyzed centromeric proteins using gene knocking-out method developed in chicken DT40 and also measured replication timing of the human 6th chromosome with MHC locus using DNA chip techniques. Then, we attempted to transfer human chromosomes from DT40 cells to mouse ES cells.
The knowledge acquired in the present study has given basic knowledge to create transgenic mice that hold the long functional genome regions such as human MHC locus. Less

  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] A.Nishihashi: "CENP-I is essential for centromere function in vertebrate cells"Developmental Cell. 2. 463-476 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.M.Spence: "Co-localisation of Centromere Activity, CEN Proteins and a Topoisomerase II Cleavage Site to a fixed region of limited extent within the Human X-satellite Locus DXZ1"The EMBO Journal. 21. 5269-5280 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Hayashi: "Ubc9 is essential for viability of higher eukaryotic cells"Experimental Cell Research. 280. 212-221 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Watanabe: "Intranuclear arrangement of human chromosome 12 correlates to large-scale replication domains"Human Molecular Genetics. 11. 13-21 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Ohno: "Triplex-forming DNAs in the human interphase nucleus visualized in situ by polypurine/polypyrimidine DNA probes and antitriplex antibodies"Chromosoma. 111. 201-213 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Kanaya: "Analysis of codon usage diversity of bacterial genes with a self-organizing map (SOM) : characterization of horizontally transferred genes with emphasis on the E. coli O157 genome"Gene. 276. 89-99 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Fukagawa: "CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells"The EMBO Journal. 20. 4603-4617 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Fukagawa: "Creation and characterization of temperature-sensitive CENP-C mutants in vertebrate cells"Nucleic Acids Research. 29. 3796-3803 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Okamura: "Gene Structure, chromosomal localization and immunolocalization of chicken centromere proteins CENP-C and ZW10"Gene. 262. 283-290 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Shina: "Genomic anatomy of a premier major histocompatibility complex paralogous region on chromosome 1q21-q22"Genome Research. 11. 789-802 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Nakamura: "Non-B DNA structures spatially and sequence-specifically associated with individual centromeres in the human interphase nucleus"Nucleic Acids Research. 28. 292 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Ohno: "Non-B DNA structures spatially and sequence-specifically associated with individua centromeres in the human interphase nucleus"Chromosomes Today. 13. 57-69 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Nogami: "Intranuclear arrangement of human chromosome 12 correlates to large-scale replication domains"Chromosoma. 108. 514-522 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Nogami: "Relative locations of the centromere and imprinted SNRPN gene within chromosome 15 territories during the cell cycle in HL6O cells"Journal of Cell Science. 113. 2157-2165 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Watanabe: "Replication timing of the human X-inactivation center (XIC) region : correlation with chromosome bands"Gene. 252. 163-172 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Kanaya: "Distribution of polypurine/polypyrimidine tract sequences in the human MHC region and their possible functions"Major Histocompatibillty Complex, Springer-Verlag Berlin Heidelberg New York Tokyo. 13 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ai Nishihashi, Tokuko Haraguchi, Yasushi Hiraoka, Toshimichi Ikemura, Vinciane Regnier, Helen Dodson, William C. Earnshaw, and Tatsuo Fukagawa: "CENP-I is essential. for centromere function in vertebrate cells"Developmental Cell. Vol. 2. 463-476 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Jenny M. Spence, Ricky Critcher, Tom A. Ebersole, Tatsuo Fukagawa, and Christine J. Farr: "Co-localisation of Centromere Activity, CEN Proteins and a Topoisomerase II Cleavage Site to a fixed region of limited extent within the Human X a-satellite Locus DXZ1"The EMBO Journal. Vol. 21. 5269-5280 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tomoko Hayashi, Masayuki Seki, Daisuke Maeda , Wensheng Wang Yoh-ichi Kawabe, Takahiko Seki, Hisato Saitoh, Tatsuo Fukagawa, Hideki Yagi, and Takemi Enomoto: "Ubc9 is essential for viability of higher eukaryotic cells"Experimental Cell Research. Vol. 280. 212-221 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mizuki Ohno, Tatsuo Fukagawa, Jeremy S. Lee, and Toshimichi Ikemura: "Triplex-forming DNAs in the human interphase nucleus visualized in situ by polypurine/polypyrimidine DNA prob es and anu' triplex antibodies"Chromosoma. Vol. 111. 201-213 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe Y, Fujiyama A, Ichiba Y, Hattori M, Yada T, Sakaki Y, Ikemura T.: "Chromosome-wide assessment of replication timing for human chromosomes 11q and 21q : disease-related genes in timing-switch regions."Human Molecular Genetics.. Vol. 11. 13-21 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tatsuo Fukagawa, Yoshikazu Mikami , Ai Nishihashi , Vinciane Regnier, Tokuko Haraguchi, Yasushi Hiraoka, Naoko Sugata, Kazuo Todokoro, William Brown, and Toshimichi Ikemura: "CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells"The EMBO Journal. Vol. 20. 4603-4617 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tatsuo Fukagawa, Vinciane Regnier, and Toshimichi Ikemura: "Creation and characterization of temperature-sensitive CENP-C mutants in vertebrate cells"Nucleic Acids Research. Vol. 29. 3796-3803 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Atsushi Okamura, Carlos Pendon, Manuel M. Valdivia, Toshimichi Ikemura, and Tatsuo Fukagawa: "Gene Structure, chromosomal localization and i mmunolocalization of chicken centromere proteins CENP-C and ZW10"Gene. VoL 262. 283-290 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shiina T, Ando A, Suto Y, Kasai F, Shigenari A, Takishima N, Kikkawa E, Iwata K, Kuwano Y, Kitamura Y, Matsuzawa Y, Sano K, Nogami M, Kawata H, Li S, Fukuzumi Y, Yamazaki M, Tashiro H, Tamiya G, Kohd A, Okumura K, Ikemura T, Soeda E, Mizuki H, Kimura M, Bahram S, Inoko H.: "Gfcnomic anatomy of a premier major histocompatibility complex paralogous region on chromosome 1q21-q22"Genome Research.. Vol. 11. 789-802 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kanaya S, Kinouchi M, Abe T, Kudo Y, Yamada Y, Nishi T, Mori H, Ikemura T.: "Analysis of codon usage diversity of bacterial genes with a self-organizing map (SOM) : characterization of horizontally transferred genes with emphasis on the E. coli O157 genome"Gene. Vol. 276. 89-99 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mizuki Ohno, Toyoaki Tenzen, Yossihisa Watanabe, Tetsushi Yamagata, Shigehiko Kanaya, Toshimichi Ikemura: "Non-B DNA structures spatially and sequence-specifically associated with individual centromeres in the human inteiphase nucleus"Chromosomes Today. Vol. 13. 57-69 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakamura Y, Gojobori T, Ikemura T.: "Codon usage tabulated from international DNA sequence databases : status for the year 2000"Nucleic Acids Res.. Vol. 28. 292 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nogami M, Nogami O, Kagotani K, Okumura M, Taguchi H, Ikemura T, Okumura K: "Intranuclear arrangement of human chromosome 12 correlates to large-scale replication domains."Chromosoma. Vol. 108. 514-522 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nogami M, Kohda A, Taguchi H, Nakao M, Ikemura T, Okumura K.: "Relative locations of the centromere and imprinted SNRPN gene within chromosome 15 territories during the cell cycle in HL60 cells"Journal of Cell Science.. Vol. 113. 2157-2165 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Watanabe Y, Tenzen T, Nagasaka Y, Inoko H, Ikemura T.: "Replication timing of the human X-inactivation center (XIC) region : correlation with chromosome bands"Gene. Vol. 252. 163-172 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2004-04-14  

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