Gene Therapy for Parkinson’s Disease - Viral Vector Approaches in Neurobiology and Brain Diseases - page 180

Fig. 3 Effi cient, regulated, and specifi c expression of lentiviral vectors using a combination of promoter and
miRNA targeting. Lentiviral vectors expressing GFP under the GFAP promoter and harboring mi124T sites were
injected into the right striatum of Sprague-Dawley rats. Three weeks after transduction, the right striatum was
lesioned with ibotenic acid (ibo). One week after ibo lesion, the animals were euthanized. The brains were
processed for histology and immunofl uorescence for GFP and GFAP was used to determine the effi ciency of
astrocytic transduction ( a ). The GFAP-GFP-mi124T was able to elicit effi cient astrocytic transduction.
Furthermore, analysis of autofl uorescence ( b ) indicated that the GFAP-GFP-mi124T was able to upregulate
GFP expression during astrogliosis. Scale bars 50 μ m

be used. Genomatix Eldorado ( http://www.genomatix.de )
analyzes the upstream region of genes for transcription factor
binding sites and suggests shorter promoters (around 700 bp).
This program also allows comparisons between species.

2. The possible length of the promoter will be limited by the type
of viral vector used and length of the transgene. We recom-
mend the maximal promoter length to be set to 4,000 bp
when using lentiviral vectors. This will leave room for a range
of transgenes and ensure that the plasmid is readily packaged
into the viral envelope.
3. Dopamine depletion may give a different change in gene
expression in different species and models. The vectors should
therefore be evaluated in all animal species and models where
it is going to be used. Such differences could potentially be
evaluated by comparing microarray data from patients to
microarray data from the animal model. Another more simple