Inducing primary brainstem gliomas in genetically engineered mice using RCAS/TVA retroviruses and Cre/loxP recombination.

Genetically engineered mice are commonly used to model brainstem gliomas in pre-clinical research. One technique for inducing primary tumors in these genetically engineered mice involves delivering viral vectors containing the code for gene-editing proteins. We present a protocol for generating primary brainstem gliomas using the RCAS-TVA retroviral delivery system and the Cre/loxP gene editing system. We describe steps for transfecting and harvesting chicken fibroblast cells, intracranially injecting cells into mice, imaging primary tumors, and treating primary tumors with focal, image-guided brain irradiation. For complete details on the use and execution of this protocol, please refer to Deland et al. (2021). ¹ .
Competing Interests: Declaration of interests D.G.K. is a cofounder of and stockholder in XRAD Therapeutics, which is developing radiosensitizers. D.G.K. is a member of the scientific advisory board and owns stock in Lumicell Inc, a company commercializing intraoperative imaging technology. None of these affiliations represents a conflict of interest with respect to the work described in this manuscript. D.G.K. is a coinventor on a patent for a handheld imaging device and is a coinventor on a patent for radiosensitizers. XRAD Therapeutics, Merck, Bristol Myers Squibb, and Varian Medical Systems provide research support to DGK, but this did not support the research described in this manuscript. Z.J.R. receives honoraria from Oakstone publishing and Eisai Pharmaceuticals for educational seminars. Z.J.R. receives royalties for intellectual property that is managed by Duke Office of Licensing and Ventures which has been licensed to Genetron Health. These sources did not support the research described in this manuscript.
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