Construction of Oncolytic Herpes Simplex Virus with Therapeutic Genes of Interest

  • Andranik Kahramanian
  • Toshihiko Kuroda
  • Hiroaki WakimotoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1937)


Herpes simplex virus (HSV) is one of the most extensively studied oncolytic virus platforms. The recent FDA approval of talimogene laherparepvec (T-VEC) has been accelerating translational research of oncolytic HSV (oHSV) as a promising therapeutic for refractory cancers such as glioblastoma, the deadliest primary malignancy in the brain. The large genome size of HSV readily allows arming of oHSV by incorporating therapeutic transgenes within the virus, as exemplified by T-VEC carrying GM-CSF, thereby enhancing the anticancer activity of oHSV. Here we describe a bacterial artificial chromosome-based method for construction of an oHSV expressing a transgene, which we routinely use in the laboratory to create a number of different recombinant oHSV bearing either therapeutic or reporter genes.

Key words

Herpes simplex virus (HSV) Glioblastoma Bacterial artificial chromosome Cre recombination Flp recombination 


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Copyright information

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

Authors and Affiliations

  • Andranik Kahramanian
    • 1
  • Toshihiko Kuroda
    • 1
  • Hiroaki Wakimoto
    • 1
    Email author
  1. 1.Department of Neurosurgery, Brain Tumor Research CenterMassachusetts General Hospital, Harvard Medical SchoolBostonUSA

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