PMID- 17178422
OWN - NLM
STAT- MEDLINE
DCOM- 20070612
LR  - 20191210
IS  - 0165-0270 (Print)
IS  - 0165-0270 (Linking)
VI  - 161
IP  - 2
DP  - 2007 Apr 15
TI  - Verification of functional AAV-mediated neurotrophic and anti-apoptotic factor 
      expression.
PG  - 291-300
AB  - The use of viral vectors for gene delivery offer many advantages for both basic 
      research and therapeutic application through the continuous expression of a gene 
      product within a target region. It is vital however that any gene product is 
      correctly expressed in a biologically active form, and this should be confirmed 
      prior to large scale in vivo studies. Using adeno-associated viral (AAV) vectors 
      to direct the expression of either a neurotrophic factor or an anti-apoptotic 
      protein, we have developed a range of in vitro assays to verify functional 
      transgenic protein expression. Brain-derived neurotropic factor (BDNF) activity 
      was confirmed by demonstrating enhanced generation of GABAergic neurons in 
      embryonic (E15) striatal cultures and AAV-mediated glial-derived neurotrophic 
      factor (GDNF) function using an assay for dopaminergic differentiation of 
      embryonic (E14) ventral mesencephalic cultures. To assess functional 
      anti-apoptotic factor expression we designed cell-survival assays, using 
      embryonic cortical cultures to confirm Bcl-x(L) activity and the HT1080 cell-line 
      for X-linked inhibitor of apoptosis protein (XIAP) activity following 
      AAV-mediated expression. This study demonstrates that the use of functional 
      assays provides valuable confirmation of desired biotherapeutic expression prior 
      to extensive investigation with new gene delivery vectors.
FAU - Kells, Adrian P
AU  - Kells AP
AD  - Neural Repair and Neurogenesis Laboratory, Department of Pharmacology and 
      Clinical Pharmacology, The University of Auckland, Auckland, New Zealand.
FAU - Henry, Rebecca A
AU  - Henry RA
FAU - Hughes, Stephanie M
AU  - Hughes SM
FAU - Connor, Bronwen
AU  - Connor B
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Review
PT  - Validation Study
DEP - 20061218
PL  - Netherlands
TA  - J Neurosci Methods
JT  - Journal of neuroscience methods
JID - 7905558
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Glial Cell Line-Derived Neurotrophic Factor)
RN  - 0 (Inhibitor of Apoptosis Proteins)
RN  - 0 (Nerve Growth Factors)
SB  - IM
MH  - Adenoviridae/*genetics
MH  - Apoptosis/*physiology
MH  - Biological Assay/*methods
MH  - Brain-Derived Neurotrophic Factor/genetics/metabolism
MH  - Genetic Vectors/genetics
MH  - Glial Cell Line-Derived Neurotrophic Factor/genetics/metabolism
MH  - Inhibitor of Apoptosis Proteins/genetics/*metabolism
MH  - Nerve Growth Factors/genetics/*metabolism
MH  - Transfection/*methods
RF  - 31
EDAT- 2006/12/21 09:00
MHDA- 2007/06/15 09:00
CRDT- 2006/12/21 09:00
PHST- 2006/08/17 00:00 [received]
PHST- 2006/11/09 00:00 [revised]
PHST- 2006/11/10 00:00 [accepted]
PHST- 2006/12/21 09:00 [pubmed]
PHST- 2007/06/15 09:00 [medline]
PHST- 2006/12/21 09:00 [entrez]
AID - S0165-0270(06)00564-4 [pii]
AID - 10.1016/j.jneumeth.2006.11.006 [doi]
PST - ppublish
SO  - J Neurosci Methods. 2007 Apr 15;161(2):291-300. doi: 
      10.1016/j.jneumeth.2006.11.006. Epub 2006 Dec 18.