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Applied Biochemistry and Biotechnology

, Volume 180, Issue 8, pp 1559–1573 | Cite as

Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting

  • Junko MasudaEmail author
  • Hiroshi Kawamoto
  • Warren Strober
  • Eiji Takayama
  • Akifumi Mizutani
  • Hiroshi Murakami
  • Tomokatsu Ikawa
  • Atsushi Kitani
  • Narumi Maeno
  • Tsukasa Shigehiro
  • Ayano Satoh
  • Akimasa Seno
  • Vaidyanath Arun
  • Tomonari Kasai
  • Ivan J. Fuss
  • Yoshimoto Katsura
  • Masaharu Seno
Article

Abstract

Transplantation of hematopoietic stem and progenitor cells (HSCs) i.e., self-renewing cells that retain multipotentiality, is now a widely performed therapy for many hematopoietic diseases. However, these cells are present in low number and are subject to replicative senescence after extraction; thus, the acquisition of sufficient numbers of cells for transplantation requires donors able to provide repetitive blood samples and/or methods of expanding cell numbers without disturbing cell multipotentiality. Previous studies have shown that HSCs maintain their multipotentiality and self-renewal activity if TCF3 transcription function is blocked under B cell differentiating conditions. Taking advantage of this finding to devise a new approach to HSC expansion in vitro, we constructed an episomal expression vector that specifically targets and transiently represses the TCF3 gene. This consisted of a vector encoding a transcription activator-like effector (TALE) fused to a Krüppel-associated box (KRAB) repressor. We showed that this TALE-KRAB vector repressed expression of an exogenous reporter gene in HEK293 and COS-7 cell lines and, more importantly, efficiently repressed endogenous TCF3 in a human B lymphoma cell line. These findings suggest that this vector can be used to maintain multipotentiality in HSC being subjected to a long-term expansion regimen prior to transplantation.

Keywords

TCF3 (E2A) Artificial transcription factor TALE technology 

Notes

Acknowledgments

This work was supported in part by Japan Society for the Promotion of Science (JSPS) KAKENHI, grant numbers 25860797, and Okayama Foundation for Science and Technology.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no financial or commercial conflicts of interest.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Junko Masuda
    • 1
    • 2
    Email author
  • Hiroshi Kawamoto
    • 3
    • 4
  • Warren Strober
    • 2
  • Eiji Takayama
    • 5
  • Akifumi Mizutani
    • 1
  • Hiroshi Murakami
    • 1
  • Tomokatsu Ikawa
    • 3
    • 6
  • Atsushi Kitani
    • 2
  • Narumi Maeno
    • 1
  • Tsukasa Shigehiro
    • 1
  • Ayano Satoh
    • 1
  • Akimasa Seno
    • 1
  • Vaidyanath Arun
    • 1
  • Tomonari Kasai
    • 1
  • Ivan J. Fuss
    • 2
  • Yoshimoto Katsura
    • 3
    • 7
  • Masaharu Seno
    • 1
  1. 1.Division of Medical Bioengineering, Graduate School of Natural Science and TechnologyOkayama UniversityOkayamaJapan
  2. 2.Mucosal Immunity Section, Laboratory of Host Defenses, National Institute of Allergy and Infectious DiseasesNational Institutes of HealthBethesdaUSA
  3. 3.Laboratory for Lymphocyte DevelopmentRIKEN Research Center for Allergy and ImmunologyYokohamaJapan
  4. 4.Department of Immunology, Institute for Frontier Medical SciencesKyoto UniversityKyotoJapan
  5. 5.Department of Oral BiochemistryAsahi University School of DentistryGifuJapan
  6. 6.Laboratory for Immune RegenerationRIKEN Center for Integrative Medical SciencesYokohamaJapan
  7. 7.Division of Cell Regeneration and Transplantation, Advanced Medical Research Center, School of MedicineNihon UniversityTokyoJapan

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