Oyibo, Hassana K., Znamenskiy, Petr, Oviedo, Hysell V., Zador, Anthony M. (September 2014) Long-term Cre-mediated retrograde tagging of neurons using a novel recombinant pseudorabies virus. Frontiers in Neuroanatomy, 8 (SEP). Art. no.86. ISSN 16625129
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Abstract
Brain regions contain diverse populations of neurons that project to different long-range targets. The study of these subpopulations in circuit function and behavior requires a toolkit to characterize and manipulate their activity in vivo. We have developed a novel set of reagents based on Pseudorabies Virus (PRV) for efficient and long-term genetic tagging of neurons based on their projection targets. By deleting IE180, the master transcriptional regulator in the PRV genome, we have produced a mutant virus capable of infection and transgene expression in neurons but unable to replicate in or spread from those neurons. IE180-null mutants showed no cytotoxicity, and infected neurons exhibited normal physiological function more than 45 days after infection, indicating the utility of these engineered viruses for chronic experiments. To enable rapid and convenient construction of novel IE180-null recombinants, we engineered a bacterial artificial chromosome (BAC) shuttle-vector system for moving new constructs into the PRV IE180-null genome. Using this system we generated an IE180-null recombinant virus expressing the site-specific recombinase Cre. This Cre-expressing virus (PRV-hSyn-Cre) efficiently and robustly infects neurons in vivo and activates transgene expression from Cre-dependent vectors in local and retrograde projecting populations of neurons in the mouse. We also generated an assortment of recombinant viruses expressing fluorescent proteins (mCherry, EGFP, ECFP). These viruses exhibit long-term labeling of neurons in vitro but transient labeling in vivo. Together these novel IE180-null PRV reagents expand the toolkit for targeted gene expression in the brain, facilitating functional dissection of neuronal circuits in vivo.
| Item Type: | Paper |
|---|---|
| Subjects: | organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > neuronal circuits organism description > virus Investigative techniques and equipment > Whole Brain Circuit Mapping |
| CSHL Authors: | |
| Communities: | CSHL labs > Zador lab School of Biological Sciences > Publications |
| Depositing User: | Matt Covey |
| Date: | 3 September 2014 |
| Date Deposited: | 25 Sep 2014 14:34 |
| Last Modified: | 29 Sep 2014 15:05 |
| PMCID: | PMC4153299 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/30819 |
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