Glioblastoma Induction Using SB Mediated Transposition: A Technique for Generating Novel Mouse Model Using Transposon Mediated Integration of SB Plasmid into Neonatal Mouse Brain

Abstract

Source: Calinescu, A. A. et al. Transposon Mediated Integration of Plasmid DNA into the Subventricular Zone of Neonatal Mice to Generate Novel Models of Glioblastoma. J. Vis. Exp. (2015)

This video describes an efficient technique to induce glioblastomas (GBM) using transposon DNA injected into the ventricles of neonatal mice. Cells of the subventricular zone, which take up the plasmid, transform, proliferate, and generate tumors with histopathological characteristics of human GBM.

Protocol

1. Intra-ventricular Neonatal Injections Three to four weeks prior to experiment, set up a mouse breeding cage with one male and one female mouse, and a plastic colored igloo. This provides an enriched environment and aids in the mating process. When pregnancy is confirmed, remove the male to a new cage. Eighteen days after mating, monitor cage daily for delivery. Perform intraventricular injections on postnatal day 1(P1). Preparation of the injection solution. &#82…

Representative Results

Figure 1: Experimental setup and guides for intra-ventricular injections in neonatal mice. (a) A stereotaxic frame (2) with micrometer dials is fitted with an automatic injector holding a 10 µl syringe (4). Inside the U frame, a neonatal adaptor frame is securely fastened (3). The control panel of the automatic injector (1) allows for precise selection of syringe, volum…

Materials

Stoelting's Lab Standard with Rat and Mouse Adaptors	Stoelting	51670	Other companies produce similar frames, any of them with small mouse adaptors are suitable
Quintessential Stereotaxic Injector (QSI)	Stoelting	53311	Injections can be made without it, ,but the automatic injector allows for increased reproducibility and convenience
10 μL 700 series hand fitted MICROLITER syringe	Hamilton	Model 701	This syringe model will deliver from 0.5 to 500 μL of solution reliably. Other syringes (for example 5  μL , Model 75) may also be used
30 gauge hypodermic needle (1.25", 15^{o} bevel)	Hamilton	S/O# 197462	This needle is ideal to pierce the skin and the skull of a neonatal mouse without the need of other invasive procedures. If it gets dull (doesn't easily enter the skin), it needs to be replaced.
In vivo jet PEI	Polyplus Transfection	201-10G	Aliquot in small volumes and keep at -20ºC
D-Luciferin, Potassium Salt	Goldbio.com	LuckK-1g	Other sources of firefly lucifierase are just as adequate
Hematoxylin Solution, Harris Modified	Sigma Aldrich	HHS128-4L