Saporin suicide gene therapy

Natasa Zarovni, Riccardo Vago, Maria Serena Fabbrini

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

New genes useful in suicide gene therapy are those encoding toxins such as plant ribosome-inactivating proteins (RIPs), which can irreversibly block protein synthesis, triggering apoptotic cell death. Plasmids expressing a cytosolic saporin (SAP) gene from common soapwort (Saponaria officinalis) are generated by placing the region encoding the mature plant toxin under the control of strong viral promoters and may be placed under tumor-specific promoters. The ability of the resulting constructs to inhibit protein synthesis is tested in cultured tumor cells co-transfected with a luciferase reporter gene. SAP expression driven by the cytomegalovirus (CMV) promoter (pCI-SAP) demonstrates that only 10 ng of plasmid DNA per 1.6 × 10 4 B16 melanoma cells drastically reduces luciferase reporter activity to 18% of that in control cells (1). Direct intratumoral injections are performed in an aggressive melanoma model. B16 melanoma-bearing mice injected with pCI-SAP complexed with lipofectamine or N-(2,3-dioleoyloxy-1-propyl) trimethylammonium methyl sulfate (DOTAP) show a noteworthy attenuation in tumor growth, and this effect is significantly augmented by repeated administrations of the DNA complexes. Here, we describe in detail this cost-effective and safe suicide gene approach.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages261-283
Number of pages23
Volume542
DOIs
Publication statusPublished - 2009

Publication series

NameMethods in Molecular Biology
Volume542
ISSN (Print)10643745

Keywords

  • DOTAP
  • melanoma
  • nonviral vectors
  • polyethylenimine (PEI)
  • ribosome- inactivating proteins
  • ricin
  • saporin
  • suicide gene therapy

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Saporin suicide gene therapy'. Together they form a unique fingerprint.

  • Cite this

    Zarovni, N., Vago, R., & Fabbrini, M. S. (2009). Saporin suicide gene therapy. In Methods in Molecular Biology (Vol. 542, pp. 261-283). (Methods in Molecular Biology; Vol. 542). https://doi.org/10.1007/978-1-59745-561-9_14