Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting

Junko Masuda, 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

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)1-15
Number of pages15
JournalApplied Biochemistry and Biotechnology
DOIs
Publication statusAccepted/In press - Jul 12 2016

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Keywords

  • Artificial transcription factor
  • TALE technology
  • TCF3 (E2A)

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

Masuda, J., Kawamoto, H., Strober, W., Takayama, E., Mizutani, A., Murakami, H., Ikawa, T., Kitani, A., Maeno, N., Shigehiro, T., Satoh, A., Seno, A., Arun, V., Kasai, T., Fuss, I. J., Katsura, Y., & Seno, M. (Accepted/In press). Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting. Applied Biochemistry and Biotechnology, 1-15. https://doi.org/10.1007/s12010-016-2187-4