Abstract
A number of gene delivery systems are currently being developed for potential use in gene therapy. Here, we demonstrate the feasibility of 21ΔqHAC, a newly developed human artificial chromosome (HAC), as a gene delivery system. We first introduced a 21ΔqHAC carrying an EGFP reporter gene and a geneticin-resistant gene (EGFP-21ΔqHAC) into hematopoietic cells by microcell-mediated chromosome transfer. These HAC-containing hematopoietic cells showed resistance to geneticin, expressed EGFP and retained the ability to differentiate into various lineages, and the EGFP-21ΔqHAC was successfully transduced into primary hematopoietic cells. Hematopoietic cells harboring the EGFP-21ΔqHAC could still be detected at two weeks post-transplantation in immunodeficient mice. We also showed effective expansion of hematopoietic cells by introducing the 21ΔqHAC containing ScFvg, a gp130-based chimeric receptor that transmits growth signals in response to specific-antigen of this receptor. All of these results demonstrate the usefulness of HAC in gene therapy.
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- EGFP:
-
Enhanced green fluorescein protein
- HAC:
-
Human artificial chromosome
- MMCT:
-
Microcell-mediated chromosome transfer
- hBM MNCs:
-
Human bone marrow mononuclear cells
- hCB:
-
Human cord blood
References
Doherty AM, Fisher EM (2003) Microcell-mediated chromosome transfer (MMCT): small cells with huge potential. Mamm Genome 14:583–592
Grimes BR, Warburton PE, Farr CJ (2002) Chromosome engineering: prospects for gene therapy. Gene Ther 9:713–718
Kakeda M, Hiratsuka M, Nagata K, Kuroiwa Y, Kakitani M, Katoh M, Oshimura M, Tomizuka K (2005) Human artificial chromosome (HAC) vector provides long-term therapeutic transgene expression in normal human primary fibroblasts. Gene Ther 12:852–856
Katoh M, Ayabe F, Norikane S, Okada T, Masumoto H, Horike S, Shirayoshi Y, Oshimura M (2004) Construction of a novel human artificial chromosome vector for gene delivery. Biochem Biophys Res Commun 321:280–290
Kawahara M, Kimura H, Ueda H, Nagamune T (2004) Selection of genetically modified cell population using hapten-specific antibody/receptor chimera. Biochem Biophys Res Commun 315:132–138
Kimura T, Wang J, Minamiguchi H, Fujiki H, Harada S, Okuda K, Kaneko H, Yokota S, Yasukawa K, Abe T, Sonoda Y (2000) Signal through gp130 activated by soluble interleukin (IL)-6 receptor (R) and IL-6 or IL-6R/IL-6 fusion protein enhances ex vivo expansion of human peripheral blood-derived hematopoietic progenitors. Stem Cells 18:444–452
Kuroiwa Y, Tomizuka K, Shinohara T, Kazuki Y, Yoshida H, Ohguma A, Yamamoto T, Tanaka S, Oshimura M, Ishida I (2000) Manipulation of human minichromosomes to carry greater than megabase-sized chromosome inserts. Nat Biotechnol 18:1086–1090
Larin Z, Mejia JE (2002) Advance in human artificial chromosome technology. Trends Genet 18:313–319
Ren X, Katoh M, Hoshiya H, Kurimasa A, Inoue T, Ayabe F, Shibata K, Toguchida J, Oshimura M (2005) A novel human artificial chromosome vector provides effective cell lineage-specific transgene expression in human mesenchymal stem cells. Stem Cells (in press)
Saffery R, Choo KH (2002) Strategies for engineering human chromosomes with therapeutic potential. J Gene Med 4:5–13
Ueda T, Tsuji K, Yoshino H, Ebihara Y, Yagasaki H, Hisakawa H, Mitsui T, Manabe A, Tanaka R, Kobayashi K, Ito M, Yasukawa K, Nakahata T (2000) Expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin, IL-6, and soluble IL-6 receptor. J Clin Invest 105:1013–1021
Acknowledgments
We thank Drs. K. Tomizuka, M. Kakeda, and M. Hiratsuka for valuable discussions. This study was supported in part by a Health and Labour Sciences Research Grant for Research on Human Genome, Tissue Engineering from the Ministry of Health, Labour and Welfare, Japan and by the Twenty-First Century COE Program of the Ministry of Education, Culture, Sports, Science and Technology of Japan: The Research Core for Chromosome Engineering Technology.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yamada, H., Kunisato, A., Kawahara, M. et al. Exogenous gene expression and growth regulation of hematopoietic cells via a novel human artificial chromosome. J Hum Genet 51, 147–150 (2006). https://doi.org/10.1007/s10038-005-0334-9
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1007/s10038-005-0334-9
Keywords
This article is cited by
-
A pathway from chromosome transfer to engineering resulting in human and mouse artificial chromosomes for a variety of applications to bio-medical challenges
Chromosome Research (2015)
-
De novo formed satellite DNA-based mammalian artificial chromosomes and their possible applications
Chromosome Research (2015)
-
A new generation of human artificial chromosomes for functional genomics and gene therapy
Cellular and Molecular Life Sciences (2013)
-
Advances in High-capacity Extrachromosomal Vector Technology: Episomal Maintenance, Vector Delivery, and Transgene Expression
Molecular Therapy (2008)
-
Input DNA Ratio Determines Copy Number of The 33 kb Factor IX Gene on De Novo Human Artificial Chromosomes
Molecular Therapy (2008)