Cell to cell transfer of the chromatin-packaged human β-globin gene cluster
Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced genes packaged into chromatin into target cells using a human artificial chromosome (HAC) and analyzed regulation of gene expression. The hu...
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Published in | Nucleic acids research Vol. 38; no. 5; p. e33 |
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Main Authors | , , , , |
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Language | English |
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01.03.2010
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Abstract | Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced genes packaged into chromatin into target cells using a human artificial chromosome (HAC) and analyzed regulation of gene expression. The human β-globin gene cluster was built into an HAC (globin-HAC) and introduced into mouse embryonic stem (ES) cells using microcell-mediated chromosome transfer (MMCT); the adult-type human β-globin gene was expressed in bone marrow and spleen cells of the transgenic mice. In vitro differentiation of ES cells into mouse erythrocytes indicated that the natural sequential expression of ε, γ and β-globin genes was reproduced on the globin-HAC. Combination of MMCT and a novel chromosome transfection technique allowed transfer of globin-HAC from HT1080 cells into the human leukemia cell line K562, and from K562 cells back into HT1080 cells. Expression of the γ-globin gene, repressed in HT1080 cells, was activated in K562 cells without any processes of differentiation into adult erythroid cells, and was completely repressed again in HT1080 cells when transferred back from K562 cells. Thus, transfer of target genes packaged into chromatin using a HAC was useful for functional analyses of gene regulation. |
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AbstractList | Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced genes packaged into chromatin into target cells using a human artificial chromosome (HAC) and analyzed regulation of gene expression. The human β-globin gene cluster was built into an HAC (globin-HAC) and introduced into mouse embryonic stem (ES) cells using microcell-mediated chromosome transfer (MMCT); the adult-type human β-globin gene was expressed in bone marrow and spleen cells of the transgenic mice. In vitro differentiation of ES cells into mouse erythrocytes indicated that the natural sequential expression of ε, γ and β-globin genes was reproduced on the globin-HAC. Combination of MMCT and a novel chromosome transfection technique allowed transfer of globin-HAC from HT1080 cells into the human leukemia cell line K562, and from K562 cells back into HT1080 cells. Expression of the γ-globin gene, repressed in HT1080 cells, was activated in K562 cells without any processes of differentiation into adult erythroid cells, and was completely repressed again in HT1080 cells when transferred back from K562 cells. Thus, transfer of target genes packaged into chromatin using a HAC was useful for functional analyses of gene regulation. Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced genes packaged into chromatin into target cells using a human artificial chromosome (HAC) and analyzed regulation of gene expression. The human beta-globin gene cluster was built into an HAC (globin-HAC) and introduced into mouse embryonic stem (ES) cells using microcell-mediated chromosome transfer (MMCT); the adult-type human beta-globin gene was expressed in bone marrow and spleen cells of the transgenic mice. In vitro differentiation of ES cells into mouse erythrocytes indicated that the natural sequential expression of epsilon, gamma and beta-globin genes was reproduced on the globin-HAC. Combination of MMCT and a novel chromosome transfection technique allowed transfer of globin-HAC from HT1080 cells into the human leukemia cell line K562, and from K562 cells back into HT1080 cells. Expression of the gamma-globin gene, repressed in HT1080 cells, was activated in K562 cells without any processes of differentiation into adult erythroid cells, and was completely repressed again in HT1080 cells when transferred back from K562 cells. Thus, transfer of target genes packaged into chromatin using a HAC was useful for functional analyses of gene regulation. Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced genes packaged into chromatin into target cells using a human artificial chromosome (HAC) and analyzed regulation of gene expression. The human β-globin gene cluster was built into an HAC (globin-HAC) and introduced into mouse embryonic stem (ES) cells using microcell-mediated chromosome transfer (MMCT); the adult-type human β-globin gene was expressed in bone marrow and spleen cells of the transgenic mice. In vitro differentiation of ES cells into mouse erythrocytes indicated that the natural sequential expression of ε, γ and β-globin genes was reproduced on the globin-HAC. Combination of MMCT and a novel chromosome transfection technique allowed transfer of globin-HAC from HT1080 cells into the human leukemia cell line K562, and from K562 cells back into HT1080 cells. Expression of the γ-globin gene, repressed in HT1080 cells, was activated in K562 cells without any processes of differentiation into adult erythroid cells, and was completely repressed again in HT1080 cells when transferred back from K562 cells. Thus, transfer of target genes packaged into chromatin using a HAC was useful for functional analyses of gene regulation. |
Author | Hasegawa, Yoshinori Suzuki, Nobutaka Ikeno, Masashi Okazaki, Tsuneko Itou, Toshihide |
AuthorAffiliation | 1 Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192 and 2 Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Kawaguchi, Saitama 332-0012, Japan |
AuthorAffiliation_xml | – name: 1 Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192 and 2 Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Kawaguchi, Saitama 332-0012, Japan |
Author_xml | – sequence: 1 fullname: Suzuki, Nobutaka – sequence: 2 fullname: Itou, Toshihide – sequence: 3 fullname: Hasegawa, Yoshinori – sequence: 4 fullname: Okazaki, Tsuneko – sequence: 5 fullname: Ikeno, Masashi |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/20007595$$D View this record in MEDLINE/PubMed |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present addresses:Nobutaka Suzuki, Yoshinori Hasegawa, Tsuneko Okazaki and Masashi Ikeno, Chromo Research Inc., 1326 Shihongi, Midori-ku, Nagoya, Aichi 458-0039, Japan Masashi Ikeno, School of Medicine, Keio University, Shinanomachi, Tokyo 160-8582, Japan |
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Snippet | Cell type-specific gene expression is regulated by chromatin structure and the transcription factors provided by the cells. In the present study, we introduced... |
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StartPage | e33 |
SubjectTerms | Animals beta-Globins - genetics beta-Globins - metabolism Cell Differentiation Cell Line Chromatin - genetics Chromosomes, Artificial, Human Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Erythrocytes - cytology Erythrocytes - metabolism Gene Expression Regulation Gene Transfer Techniques Humans K562 Cells Methods Online Mice Mice, Transgenic Multigene Family |
Title | Cell to cell transfer of the chromatin-packaged human β-globin gene cluster |
URI | https://www.ncbi.nlm.nih.gov/pubmed/20007595 https://search.proquest.com/docview/733525454 https://pubmed.ncbi.nlm.nih.gov/PMC2836578 |
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