Saporin suicide gene therapy

Natasa Zarovni, Riccardo Vago, Maria Serena Fabbrini

Research output: Chapter in Book/Report/Conference proceedingChapter

10 Citations (Scopus)

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

Fingerprint

Genetic Therapy
Suicide
Experimental Melanomas
Luciferases
Saponaria
Plasmids
Ribosome Inactivating Proteins
Cultured Tumor Cells
Genes
Plant Proteins
Aptitude
DNA
Cytomegalovirus
Reporter Genes
Carcinogens
Melanoma
Proteins
Cell Death
Costs and Cost Analysis
Injections

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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

Saporin suicide gene therapy. / Zarovni, Natasa; Vago, Riccardo; Fabbrini, Maria Serena.

Methods in Molecular Biology. Vol. 542 2009. p. 261-283 (Methods in Molecular Biology; Vol. 542).

Research output: Chapter in Book/Report/Conference proceedingChapter

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