Construction of Minicircle Suicide Genes Coding for Ribosome-Inactivating Proteins

Methods Mol Biol. 2022:2521:157-171. doi: 10.1007/978-1-0716-2441-8_8.

Abstract

Due to the lower risks of adverse effects, nonviral gene therapy is a suitable alternative to transfect cancer cells with a suicide gene to let them kill themselves by expressing toxic ribosome-inactivating proteins. Plasmids are stable and easy-to-produce vectors, but they have some disadvantages due to the bacterial backbone. Applying the minicircle technology, this problem can be solved with manageable effort in a well-equipped laboratory. With the described methodology, minicircle-DNA can be produced at low costs. The cell killing properties are monitored following transfection using the CytoSMART® Omni system-a camera based live cell imaging device.

Keywords: Gene delivery; Live cell imaging; Minicircle-DNA; Ribosome-inactivating protein; Suicide gene.

MeSH terms

  • Genetic Vectors* / genetics
  • Humans
  • Plasmids / genetics
  • Ribosome Inactivating Proteins*
  • Ribosomes / genetics
  • Transfection

Substances

  • Ribosome Inactivating Proteins