@article {49289,
	title = {A transgenic mouse for imaging activity-dependent dynamics of endogenous Arc mRNA in live neurons.},
	journal = {Science Advances},
	volume = {4},
	year = {2018},
	month = {2018 Jun},
	pages = {eaar3448},
	abstract = {<p>Localized translation plays a crucial role in synaptic plasticity and memory consolidation. However, it has not been possible to follow the dynamics of memory-associated mRNAs in living neurons in response to neuronal activity in real time. We have generated a novel mouse model where the endogenous Arc/Arg3.1 gene is tagged in its 3{\textquoteright} untranslated region with stem-loops that bind a bacteriophage PP7 coat protein (PCP), allowing visualization of individual mRNAs in real time. The physiological response of the tagged gene to neuronal activity is identical to endogenous Arc and reports the true dynamics of Arc mRNA from transcription to degradation. The transcription dynamics of Arc in cultured hippocampal neurons revealed two novel results: (i) A robust transcriptional burst with prolonged ON state occurs after stimulation, and (ii) transcription cycles continue even after initial stimulation is removed. The correlation of stimulation with Arc transcription and mRNA transport in individual neurons revealed that stimulus-induced Ca activity was necessary but not sufficient for triggering Arc transcription and that blocking neuronal activity did not affect the dendritic transport of newly synthesized Arc mRNAs. This mouse will provide an important reagent to investigate how individual neurons transduce activity into spatiotemporal regulation of gene expression at the synapse.</p>
},
	issn = {2375-2548},
	doi = {10.1126/sciadv.aar3448},
	author = {Das, Sulagna and Moon, Hyungseok C and Singer, Robert H and Park, Hye Yoon}
}
@article {6601,
	title = {A transgenic mouse for in vivo detection of endogenous labeled mRNA.},
	journal = {Nature Methods},
	volume = {8},
	year = {2011},
	month = {2011 Feb},
	pages = {165-70},
	abstract = {<p>Live-cell single mRNA imaging is a powerful tool but has been restricted in higher eukaryotes to artificial cell lines and reporter genes. We describe an approach that enables live-cell imaging of single endogenous labeled mRNA molecules transcribed in primary mammalian cells and tissue. We generated a knock-in mouse line with an MS2 binding site (MBS) cassette targeted to the 3{\textquoteright} untranslated region of the essential β-actin gene. As β-actin-MBS was ubiquitously expressed, we could uniquely address endogenous mRNA regulation in any tissue or cell type. We simultaneously followed transcription from the β-actin alleles in real time and observed transcriptional bursting in response to serum stimulation with precise temporal resolution. We tracked single endogenous labeled mRNA particles being transported in primary hippocampal neurons. The MBS cassette also enabled high-sensitivity fluorescence in situ hybridization (FISH), allowing detection and localization of single β-actin mRNA molecules in various mouse tissues.</p>
},
	keywords = {Animals, Cell Survival, Cells, Cultured, Fluorescence, Genetic, In Situ Hybridization, Messenger, Mice, RNA, Transcription, Transgenic},
	issn = {1548-7105},
	doi = {10.1038/nmeth.1551},
	author = {Lionnet, Timoth{\'e}e and Czaplinski, Kevin and Darzacq, Xavier and Shav-Tal, Yaron and Wells, Amber L and Chao, Jeffrey A and Park, Hye Yoon and de Turris, Valeria and Lopez-Jones, Melissa and Singer, Robert H}
}
