Functional knock-out of c-myb by an intracellular anti-c-Myb single-chain antibody

Keizo Kasono, Alain Piché, Jialing Xiang, Hyung Gyoon Kim, Guadalupe Bilbao, Feng Johanning, Michael Nawrath, Karin Moelling, David T. Curiel

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Aberrant expression of the c-myb proto-oncogene is a key factor in the development of the neoplastic phenotype in a variety of contexts. On this basis, it has been proposed that ablation of c-myb function might be an effective approach for therapy. To this end, we have employed an intracellular single-chain antibody (sFv) approach to achieve the functional knock-out of the c-Myb once-protein. We derived an anti-c-Myb sFv, which was configured into eukaryotic expression plasmids. We confirmed the expression of the cytoplasmic and nuclear forms of the sFvs in the correct subcellular compartments by immunofluorescent staining. Importantly, the anti-c-Myb sFvs strongly inhibited the transactivation activity of c-Myb. Furthermore, cytotoxic effect of the sFv was observed only in the c-Myb positive cell line K562. These results suggest that anti-c-Myb sFv is a valuable tool for understanding the molecular mechanisms of c-myb induced transformation. In addition, this approach may have potential utility in the gene therapy for c-myb dependent malignant diseases.

Original languageEnglish
Pages (from-to)124-130
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume251
Issue number1
DOIs
StatePublished - Oct 9 1998
Externally publishedYes

Keywords

  • Gene therapy
  • Leukemia
  • Single-chain antibody
  • Transactivation
  • c-myb

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    Kasono, K., Piché, A., Xiang, J., Kim, H. G., Bilbao, G., Johanning, F., Nawrath, M., Moelling, K., & Curiel, D. T. (1998). Functional knock-out of c-myb by an intracellular anti-c-Myb single-chain antibody. Biochemical and Biophysical Research Communications, 251(1), 124-130. https://doi.org/10.1006/bbrc.1998.9259