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bioRxiv · 10.1101/2022.09.19.508525

Inducible CRISPR epigenome systems mimic cocaine induced bidirectional regulation of Nab2 and Egr3

Choi, E. Y.·Franco, D.·Stapf, C. A.·Gordin, M.·Chow, A.·Cover, K. K.·Chandra, R.·Lobo, M. K.
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Abstract

Substance use disorder is a debilitating chronic disease and a leading cause of disability around the world. The nucleus accumbens (NAc) is a major brain hub that mediates reward behavior. Studies demonstrate exposure to cocaine is associated with molecular and functional imbalance in two NAc medium spiny neuron subtypes (MSNs), dopamine receptor 1 and 2 enriched D1-MSNs and D2-MSNs. Our previous reports showed that repeated cocaine exposure induced transcription factor early growth response 3 (Egr3) mRNA in NAc D1-MSNs, while reducing it in D2-MSNs. Here, we report our findings of repeated cocaine exposure inducing cell subtype specific bidirectional expression of the Egr3 corepressor NGFI-A-binding protein 2 (Nab2). Using CRISPR activation and interference (CRISPRa and CRISPRi) tools combined with Nab2 or Egr3 targeted sgRNAs, we mimicked these bidirectional changes in Neuro2a cells. Furthermore, we investigated D1-MSN and D2-MSN subtype specific expressional changes of histone lysine demethylases Kdm1a, Kdm6a and Kdm5c in NAc after repeated cocaine exposure. Since Kdm1a showed bidirectional expression patterns in D1-MSNs and D2-MSNs, like Egr3, we developed a light inducible Opto-CRISPR-KDM1a system. We were able to downregulate Egr3 and Nab2 transcripts and cause bidirectional expression changes in D1-MSNs and D2-MSNs similar to cocaine exposure in Neuro2A cells. In contrast, our Opto-CRISPR-p300 activation system induced the Egr3 and Nab2 transcripts and caused bidirectional transcription regulations in D1-MSNs and D2-MSNs. Our study sheds light on the expression patterns of Nab2 and Egr3 in specific NAc MSN subtypes in cocaine action and uses CRISPR tools to further mimic these expression patterns.

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Choi, E. Y., Franco, D., Stapf, C. A., Gordin, M., Chow, A., Cover, K. K., Chandra, R., Lobo, M. K.. 2022-09-19. Inducible CRISPR epigenome systems mimic cocaine induced bidirectional regulation of Nab2 and Egr3. https://doi.org/10.1101/2022.09.19.508525

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