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      A robust and high-throughput Cre reporting and characterization system for the whole mouse brain

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          Abstract

          The Cre/lox system is widely used in mice to achieve cell-type-specific gene expression. However, a strong and universal responding system to express genes under Cre control is still lacking. We have generated a set of Cre reporter mice with strong, ubiquitous expression of fluorescent proteins of different spectra. The robust native fluorescence of these reporters enables direct visualization of fine dendritic structures and axonal projections of the labeled neurons, which is useful in mapping neuronal circuitry, imaging and tracking specific cell populations in vivo. Using these reporters and a high-throughput in situ hybridization platform, we are systematically profiling Cre-directed gene expression throughout the mouse brain in a number of Cre-driver lines, including novel Cre lines targeting different cell types in the cortex. Our expression data are displayed in a public online database to help researchers assess the utility of various Cre-driver lines for cell-type-specific genetic manipulation.

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          Most cited references33

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          Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide-based retroviral vector.

          Attempts to generate reliable and versatile vectors for gene therapy and biomedical research that express multiple genes have met with limited success. Here we used Picornavirus 'self-cleaving' 2A peptides, or 2A-like sequences from other viruses, to generate multicistronic retroviral vectors with efficient translation of four cistrons. Using the T-cell receptor:CD3 complex as a test system, we show that a single 2A peptide-linked retroviral vector can be used to generate all four CD3 proteins (CD3epsilon, gamma, delta, zeta), and restore T-cell development and function in CD3-deficient mice. We also show complete 2A peptide-mediated 'cleavage' and stoichiometric production of two fluorescent proteins using a fluorescence resonance energy transfer-based system in multiple cell types including blood, thymus, spleen, bone marrow and early stem cell progenitors.
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            Subregion- and cell type-restricted gene knockout in mouse brain.

            Using the phage P1-derived Cre/loxP recombination system, we have developed a method to create mice in which the deletion (knockout) of virtually any gene of interest is restricted to a subregion or a specific cell type in the brain such as the pyramidal cells of the hippocampal CA1 region. The Cre/loxP recombination-based gene deletion appears to require a certain level of Cre protein expression. The brain subregional restricted gene knockout should allow a more precise analysis of the impact of a gene mutation on animal behaviors.
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              A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping.

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                Author and article information

                Journal
                9809671
                21092
                Nat Neurosci
                Nature neuroscience
                1097-6256
                1546-1726
                22 December 2009
                20 December 2009
                January 2010
                1 July 2010
                : 13
                : 1
                : 133-140
                Affiliations
                [1 ] Allen Institute for Brain Science, Seattle, WA 98103, USA
                [2 ] Howard Hughes Medical Institute, Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
                Author notes
                [* ]Corresponding author. Phone: (206) 548-7104; hongkuiz@ 123456allenstitute.org
                Article
                nihpa165655
                10.1038/nn.2467
                2840225
                20023653
                29b517cf-adb1-4167-b08f-c7901ed5ca41

                Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms

                History
                Funding
                Funded by: National Institute of Mental Health : NIMH
                Award ID: R01 MH085500-01 ||MH
                Categories
                Article

                Neurosciences
                Neurosciences

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