Abstract
ABSTRACTIt is important to study the neural connectivities and functions in primates. For this purpose, it is critical to be able to transfer genes to certain neurons in the primate brain so that we can image the neuronal signals and analyze the function of the transferred gene. Toward this end, our team has been developing gene transfer systems using viral vectors. In this review, we summarize our current achievements as follows. 1) We compared the features of gene transfer using five different AAV serotypes in combination with three different promoters, namely, CMV, mouse CaMKII (CaMKII), and human synapsin 1 (hSyn1), in the marmoset cortex with those in the mouse and macaque cortices. 2) We used target‐specific double‐infection techniques in combination with TET‐ON and TET‐OFF using lentiviral retrograde vectors for enhanced visualization of neural connections. 3) We used an AAV‐mediated gene transfer method to study the transcriptional control for amplifying fluorescent signals using the TET/TRE system in the primate neocortex. We also established systems for shRNA mediated gene targeting in a neocortical region where a gene is significantly expressed and for expressing the gene using the CMV promoter for an unexpressed neocortical area in the primate cortex using AAV vectors to understand the regulation of downstream genes. Our findings have demonstrated the feasibility of using viral vector mediated gene transfer systems for the study of primate cortical circuits using the marmoset as an animal model. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 354–372, 2017
Highlights
The neocortex has emerged in mammals during the course of the evolution of their brain
1) We compared the features of gene transfer using five different associated virus (AAV) serotypes in combination with three different promoters, namely, CMV, mouse CaMKII (CaMKII), and human synapsin 1, in the marmoset cortex with those in the mouse and macaque cortices
Our findings have demonstrated the feasibility of using viral vector mediated gene transfer systems for the study of primate cortical circuits using the marmoset as an animal model
Summary
The neocortex has emerged in mammals during the course of the evolution of their brain. In this review we focus on the topics that are directly related to our recently reported applications of viral vectors for gene transfer in the primate cortex, in the marmoset brain. There are several issues that remain to be solved in using AAV vectors They include AAV serotypes, promoters, toxicity, and species specificity (Nassi et al, 2015b; EI-Shamayleh et al, 2016). We considered that it is important to determine the factors that influence the efficacies of gene transfer and tropism To this end, we compared the infectious properties of five different capsids of AAV serotypes 1, 2, 5, 8, and 9 in marmosets as well as in mice and macaques. AAV serotypes of 2, 5, 8, and 9 can be used in nonhuman primates (Diester et al, 2011; Cavanaugh et al 2012; Jazayeri et al, 2012; Ohayon et al, 2013; Nassi et al, 2015a; Afraz et al, 2015; Inoue et al, 2015; MacDougall et al, in press; Table 1 Summary of AAV Serotpye Application in Mammalian Brains (mice)
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