Arc-TimeSTAMP2 mice have the TimeSTAMP cassette replacing the endogenous STOP codon of the activity regulated cytoskeletal-associated protein (Arc) gene. ARC is a highly conserved cytoskeleton-associated protein that is localized to the nucleus, cytosol, and postsynapse where it interacts both indirectly with the actin cytoskeleton and directly with synaptic scaffold proteins. ARC is restricted to a subset of neurons that are CaMKII-positive, GABA-negative, and glutamatergic where it plays a role in memory and synaptic plasticity. Specifically, the TimeSTAMP (Time-Specific Tagging for the Age Measurement of Proteins) cassette contains N- and C-terminal Venus yellow fluorescent protein (YFP) fragments separated by a cis-cleaving hepatitis C virus (HCV) NS3 protease, and a 2xHA-tag. The NS3 non-virulent protease will become active after the fusion protein is synthesized and will cleave the substrates N-terminal and C-terminal before the YFP halves can assemble, leaving ARC tagged with a nonfluorescent N-terminal half of YFP. However, when an NS3 inhibitor drug such as asunaprevir (ASV) is applied, the fusion proteins will not undergo cleavage, allowing for the non-reversible folding, complementation, and maturation of the YFP. This permits detection of newly synthesized protein copies by fluorescence microscopy. ARC will also be labeled by the 2xHA-tag after introduction of ASV. This TimeSTAMP2 tag remains stable even if the drug is washed out and the protease becomes active and cleaves the tag's two substrates. Furthermore, the C-terminal YFP also contains two HA epitope tags. Thus, copies of ARC synthesized during time of drug presence are selectively labeled. The donating investigator reports that homozygous and heterozygous mice are viable and fertile. This strain is useful for visualizing newly synthesized ARC protein where preservation of normal neuronal function is required.

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Arc-TimeSTAMP2 mice have the TimeSTAMP cassette replacing the endogenous STOP codon of the Arc gene. The TimeSTAMP (Time-Specific Tagging for the Age Measurement of Proteins) cassette contains N- and C-terminal Venus fragments separated by a cis-cleaving hepatitis C virus (HCV) NS3 protease, and a 2xHA-tag. Expression of Arc-TimeSTAMP is achieved only after administration of the NS3 inhibitor drug asunaprevir (ASV). This strain is useful for visualizing newly synthesized ARC where preservation of normal neuronal function is required.

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