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Neurosci."],"published-print":{"date-parts":[[2023,3,29]]},"abstract":"<jats:p>\n                    Substance use disorder is a chronic disease and a leading cause of disability around the world. The NAc is a major brain hub mediating reward behavior. Studies demonstrate exposure to cocaine is associated with molecular and functional imbalance in NAc medium spiny neuron subtypes (MSNs), dopamine receptor 1 and 2 enriched D1-MSNs and D2-MSNs. We previously reported repeated cocaine exposure induced transcription factor early growth response 3 (\n                    <jats:italic>Egr3<\/jats:italic>\n                    ) mRNA in NAc D1-MSNs, and reduced it in D2-MSNs. Here, we report our findings of repeated cocaine exposure in male mice inducing MSN subtype-specific bidirectional expression of the\n                    <jats:italic>Egr3<\/jats:italic>\n                    corepressor NGFI-A-binding protein 2 (\n                    <jats:italic>Nab2<\/jats:italic>\n                    ). Using CRISPR activation and interference (CRISPRa and CRISPRi) tools combined with\n                    <jats:italic>Nab2<\/jats:italic>\n                    or\n                    <jats:italic>Egr3<\/jats:italic>\n                    -targeted sgRNAs, we mimicked these bidirectional changes in Neuro2a cells. Furthermore, we investigated D1-MSN- and D2-MSN-specific expressional changes of histone lysine demethylases\n                    <jats:italic>Kdm1a<\/jats:italic>\n                    ,\n                    <jats:italic>Kdm6a<\/jats:italic>\n                    , and\n                    <jats:italic>Kdm5c<\/jats:italic>\n                    in NAc after repeated cocaine exposure in male mice. Since\n                    <jats:italic>Kdm1a<\/jats:italic>\n                    showed bidirectional expression patterns in D1-MSNs and D2-MSNs, like\n                    <jats:italic>Egr3<\/jats:italic>\n                    , we developed a light-inducible Opto-CRISPR-KDM1a system. We were able to downregulate\n                    <jats:italic>Egr3<\/jats:italic>\n                    and\n                    <jats:italic>Nab2<\/jats:italic>\n                    transcripts in Neuro2A cells and cause similar bidirectional expression changes we observed in D1-MSNs and D2-MSNs of mouse repeated cocaine exposure model. Contrastingly, our Opto-CRISPR-p300 activation system induced the\n                    <jats:italic>Egr3<\/jats:italic>\n                    and\n                    <jats:italic>Nab2<\/jats:italic>\n                    transcripts and caused opposite bidirectional transcription regulations. Our study sheds light on the expression patterns of\n                    <jats:italic>Nab2<\/jats:italic>\n                    and\n                    <jats:italic>Egr3<\/jats:italic>\n                    in specific NAc MSNs in cocaine action and uses CRISPR tools to further mimic these expression patterns.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>SIGNIFICANCE STATEMENT<\/jats:bold>\n                    Substance use disorder is a major societal issue. The lack of medication to treat cocaine addiction desperately calls for a treatment development based on precise understanding of molecular mechanisms underlying cocaine addiction. In this study, we show that\n                    <jats:italic>Egr3<\/jats:italic>\n                    and\n                    <jats:italic>Nab2<\/jats:italic>\n                    are bidirectionally regulated in mouse NAc D1-MSNs and D2-MSNs after repeated exposure to cocaine. Furthermore, histone lysine demethylations enzymes with putative EGR3 binding sites showed bidirectional regulation in D1- and D2-MSNs after repeated exposure to cocaine. Using Cre- and light-inducible CRISPR tools, we show that we can mimic this bidirectional regulation of\n                    <jats:italic>Egr3<\/jats:italic>\n                    and\n                    <jats:italic>Nab2<\/jats:italic>\n                    in Neuro2a cells.\n                  <\/jats:p>","DOI":"10.1523\/jneurosci.1802-22.2022","type":"journal-article","created":{"date-parts":[[2023,2,27]],"date-time":"2023-02-27T13:50:19Z","timestamp":1677505819000},"page":"2242-2259","source":"Crossref","is-referenced-by-count":11,"title":["Inducible CRISPR Epigenome Systems Mimic Cocaine Induced Bidirectional Regulation of Nab2 and Egr3"],"prefix":"10.1523","volume":"43","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0023-6844","authenticated-orcid":false,"given":"Eric 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