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NEURON‐SPECIFIC TRANSGENE EXPRESSION OF IMP2 MEDIATED BY SYNAPSIN PROMOTER‐DRIVEN AAV9
Author(s) -
Park Danielle,
Blizard Sarah,
Austin Scarlett,
Harman Joshua,
Haraszti Samantha,
Xu Mei
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.744.4
Subject(s) - biology , transgene , microbiology and biotechnology , axon , synapsin i , dorsal root ganglion , gene , neuroscience , genetics , spinal cord , vesicle , membrane , synaptic vesicle
Insulin‐ like growth factor‐II (IGF‐II) mRNA‐binding protein‐2 (IMP2) is one of the three homologues (IMP1‐3) that play an important role in the post‐transcriptional regulation of gene expression during development in several tissues including the nervous system. An alternative splice product of IMP2 lacking exon 10 aberrantly expresses in human hepatocellular carcinoma and is identified as HCC. Several lines of studies have demonstrated that IMP1/ZBP1 (zipcode binding protein) is critical in axon guidance and regeneration by regulating localization and translation of specific mRNAs. However, a role of IMP2 in the neural tissue is largely unknown. Previously, we used a custom made IMP2‐specific antibody to locate the endogenous IMP2 expression in mouse neurons and glial cells at different developmental stages. Our pilot study shows that IMP2 expression is sustained throughout life and it may play a role to facilitate axon regeneration. In this study, we used the synapsin promoter‐driven adeno‐associated viral (AAV) 9 constructs to express YFP‐IMP2 and –HCC specifically in neurons both in vitro and in vivo . We applied AAV9.hSyn.YFP.PolyA, AAV9.hSyn.YFP‐hIMP2.PolyA and AAV9.hSyn.YFP‐hHCC.PolyA in the primary dorsal root ganglion mouse neuron culture as an in vitro model. We also injected the viral vectors into the crushed mouse sciatic nerve as an in vivo model. Our results demonstrate the moderate neuron‐specific transgene expression of IMP2/HCC in both models. The data indicate that the synapsin promoter‐driven AAV9 can be used as an important tool for further study of the role of IMP2 in the nervous system. Support or Funding Information This work is supported by the Department of Bio‐Medical Sciences, PCOM and Health Research Grant, Pennsylvania Department of Health.