Identification of cell-type-specific promoters within the brain using lentiviral vectors

• Type of Material: Article
• Authors: JP Chhatwal, Emory University; SE Hammack, Emory University; AM Jasnow, Emory University; Donald Rainnie, Emory University; Kerry Ressler, Emory University
• Language: English
• Date: 2007-04
• Publisher: Springer Nature [academic journals on nature.com]: Hybrid Journals
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2007 Nature Publishing Group All rights reserved
• Final Published Version (URL): http://www.nature.com/gt/journal/v14/n7/full/3302898a.html
• Title of Journal or Parent Work: Gene Therapy
• ISSN: 0969-7128
• Volume: 14
• Issue: 7
• Start Page: 575
• End Page: 583
• Grant/Funding Information: National Institute on Drug Abuse : NIDA; Support was provided by NIH (MH074097, MH069884, MH070218), NARSAD, the Burrough’s Wellcome Fund, the Center for Behavioral Neuroscience (NSF agreement IBN-987675), and by NIH/NCRR base grant (P51RR000165) to Yerkes National Primate Research Center.
• Keywords: electrophysiology; interneuron; lentivirus; neuroscience; behavior; inhibitor; psychiatry
• Research Categories: Biology, Neuroscience; Psychology, Behavioral

Abstract

The development of cell-type-specific mini-promoters for genetic studies is complicated by a number of issues. Here, we describe a general method for the relatively rapid screening of specific promoter activity in cell culture, in acute brain slice preparations and in vivo. Specifically, we examine the activity of an ~3 kb promoter region from the neuroactive peptide cholecystokinin (CCK) compared to the commonly used cytomegalovirus promoter. We find a high degree of cell-type selectivity in vivo using lentiviral approaches in rats and traditional transgenic approaches in mice. Appropriate colocalization of Cre-recombinase and CCK gene expression is found within the hippocampus, when the CCK promoter is driving either the expression of Cre-recombinase or green fluorescent protein. We also demonstrate fluorescent identification of CCK-positive inter-neurons that allows for cell-type-specific electrophysiologic studies in rats and mice. In conclusion, these studies identify a functional mini-promoter for the CCK gene and outline a novel and sensitive general method to test activity of selective promoters in vitro and in vivo. This approach may allow for the more rapid identification of specific promoters for use with transgenic animals, in genetically modified viruses, and in the design of targeted, therapeutic gene-delivery systems.