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Production of Viral Vectors with Suicide Genes by Utilizing the Intron-Splicing Mechanism of Insect Cells

  • Haifeng Chen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1895)

Abstract

Viral vectors carrying suicide genes such as diphtheria toxin, Pseudomonas exotoxin, or barnase are very useful tools in cancer gene therapy and cell ablation research. However, such viral vectors are extremely difficult to produce due to the fact that trace amounts of the toxin will kill any cells used for viral vector production. To overcome this obstacle, we inserted mammalian introns that are not recognized by insect cells to break up the open reading frames (ORFs) of the toxic genes and successfully produced at normal levels of baculoviral and adeno-associated viral (AAV) vectors carrying these toxic genes. Once these viral vectors were used to infect mammalian cells, the introns were spliced out and toxic proteins expressed to kill the target cells.

Key words

Adeno-associated virus Baculovirus Insect cells Toxin Suicide genes Cancer therapy Cell ablation Purification Intron-splicing 

Notes

Acknowledgments

Min Chen gave valuable comments on the manuscript. Experiments of AAV vector production, purification, and SDS gel electrophoresis were performed by Ching Yi Ho.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Haifeng Chen
    • 1
  1. 1.R&D, Virovek, Inc.HaywardUSA

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