Gene transfer into rat brain using adenoviral vectors

PUNTEL; KROEGER; SANDERSON; THOMAS; CASTRO; LOWENSTEIN

Current protocols in Neuroscience

John Wiley & Sons, Inc.

Año: 2010

1934-8584

Viral vector–mediated gene delivery is an attractive procedure for introducing genesinto the brain, both for purposes of basic neuroscience research and to develop genetherapy for neurological diseases. Replication-defective adenoviruses possess many fea-tures which make them ideal vectors for this purpose—efÞciently transducing terminallydifferentiated cells such as neurons and glial cells, resulting in high levels of transgeneexpression in vivo. Also, in the absence of anti-adenovirus immunity, these vectors cansustain very long-term transgene expression within the brain parenchyma. This unitprovides protocols for the stereotactic injection of adenoviral vectors into the brain,followed by protocols to detect transgene expression or inÞltrates of immune cells byimmunocytochemistry or immunoßuorescence. ELISPOT and neutralizing antibody as-say methodologies are provided to quantitate the levels of cellular and humoral immuneresponses against adenoviruses. Quantitation of adenoviral vector genomes within the ratbrain using qPCR is also described. C