Generation of adeno-associated virus vector enabling functional expression of oxytocin receptor and fluorescence marker genes using the human eIF4G internal ribosome entry site element.
We developed the AAV-Oxtr-IRES-Venus vector to rescue the oxytocin receptor (Oxtr) gene functionally at restricted regions in the brains of Oxtr knockout mice. First we chose human eIF4G gene-derived IRES to co-express Venus, a fluorescent marker gene, with Oxtr. With selected human eIF4G IRES, we constructed the AAV-Oxtr-IRES-Venus vector, and it caused expression of the Venus gene in the brain when 1 microl of viral solution (9.4x10(7) vg) was injected into the medial amygdaloid nucleus. In primary neuronal cells transduced with this viral vector and followed by oxytocin administration, functional expression of OXTR was detected by Ca(2+) imaging assay.
Sato
Keisuke
K
Laboratory of Molecular Biology, Department of Molecular and Cell Biology, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, Miyagi 981-8555, Japan.
Date
Shiori
S
Aoyagi
Yumi
Y
Kasahara
Yoshiyuki
Y
Nawa
Akihiko
A
Mizukami
Hiroaki
H
Hidema
Shizu
S
Ozawa
Keiya
K
Nishimori
Katsuhiko
K
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D000229:Dependovirus / Q000235:genetics*; D039603:Eukaryotic Initiation Factor-4G / Q000235:genetics*; D005453:Fluorescence; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D018045:Receptors, Oxytocin / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism*
D017931:DNA Primers; D039603:Eukaryotic Initiation Factor-4G; D018045:Receptors, Oxytocin