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.
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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.
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Masuda, J., Kawamoto, H., Strober, W. et al. Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting. Appl Biochem Biotechnol 180, 1559–1573 (2016). https://doi.org/10.1007/s12010-016-2187-4
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DOI: https://doi.org/10.1007/s12010-016-2187-4