Glioblastoma Induction Using SB Mediated Transposition: A Technique for Generating Novel Mouse Model Using Transposon Mediated Integration of SB Plasmid into Neonatal Mouse Brain
Trascrizione
Begin by taking a tube containing a mix of plasmids. The sleeping beauty or SB transposon plasmid is engineered to carry a transposable element comprising a human-derived glioblastoma-inducing oncogene sandwiched between two inverted repeat sequences.In addition, this mix contains an SB transposase plasmid that has a transposase enzyme expressing element.
Now, add a suitable transfection reagent to the tube. Aspirate the injection mix into a syringe fitted with a microneedle. Next, secure an anesthetized mouse pup in the prone position on a stereotaxic frame. Visualize the pup's head region to locate the cranial sutures. Position the microneedle above the cranial sutures.
Inject the mix into the lateral ventricle – a cavity within the brain parenchyma. Subsequently, the solution gets dispersed into the brain parenchyma, where the transfection reagent molecules facilitate the entry of plasmids within the cells. Once inside the cell, the transposase plasmid expresses the transposase enzyme molecules, which recognize the inverted repeat sequences and excise the transposon element.
The transposable element DNA then reintegrates itself into the TA repeat region of the pup's genomic DNA. Genetic transposition results in an oncogenic transformation of brain cells, leading to the development of glioblastoma in the mouse brain.
