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A neuron-optimized CRISPR/dCas9 activation system for robust and specific gene regulation

View ORCID ProfileKatherine E. Savell, Svitlana V. Bach, Morgan E. Zipperly, Jasmin S. Revanna, Nicholas A. Goska, Jennifer J. Tuscher, Corey G. Duke, Faraz A. Sultan, Julia N. Burke, Derek M. Williams, Lara Ianov, View ORCID ProfileJeremy J Day
doi: https://doi.org/10.1101/371500
Katherine E. Savell
University of Alabama at Birmingham
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  • ORCID record for Katherine E. Savell
Svitlana V. Bach
University of Alabama at Birmingham
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Morgan E. Zipperly
University of Alabama at Birmingham
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Jasmin S. Revanna
University of Alabama at Birmingham
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Nicholas A. Goska
University of Alabama at Birmingham
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Jennifer J. Tuscher
University of Alabama at Birmingham
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Corey G. Duke
University of Alabama at Birmingham
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Faraz A. Sultan
University of Alabama at Birmingham
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Julia N. Burke
University of Alabama at Birmingham
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Derek M. Williams
University of Alabama at Birmingham
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Lara Ianov
University of Alabama at Birmingham
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Jeremy J Day
University of Alabama at Birmingham
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  • ORCID record for Jeremy J Day
  • For correspondence: jjday@uab.edu
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Abstract

Recent developments in CRISPR-based gene editing have provided new avenues to interrogate gene function. However, application of these tools in the central nervous system has been delayed due to difficulties in transgene expression in post-mitotic neurons. Here, we present a highly efficient, neuron-optimized dual lentiviral CRISPR-based transcriptional activation (CRISPRa) system to drive gene expression in primary neuronal cultures and the adult brain of rodent model systems. We demonstrate robust, modular, and tunable induction of endogenous target genes as well as multiplexed gene regulation necessary for investigation of complex transcriptional programs. CRISPRa targeting unique promoters in the complex multi-transcript gene Brain-derived neurotrophic factor (Bdnf) revealed both transcript- and genome-level selectivity of this approach, in addition to highlighting downstream transcriptional and physiological consequences of Bdnf regulation. Finally, we illustrate that CRISPRa is highly efficient in vivo, resulting in increased protein levels of a target gene in diverse brain structures. Taken together, these results demonstrate that CRISPRa is an efficient and selective method to study gene expression programs in brain health and disease.

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The copyright holder for this preprint is the author/funder. It is made available under a CC-BY-ND 4.0 International license.
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  • Posted July 17, 2018.

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A neuron-optimized CRISPR/dCas9 activation system for robust and specific gene regulation
Katherine E. Savell, Svitlana V. Bach, Morgan E. Zipperly, Jasmin S. Revanna, Nicholas A. Goska, Jennifer J. Tuscher, Corey G. Duke, Faraz A. Sultan, Julia N. Burke, Derek M. Williams, Lara Ianov, Jeremy J Day
bioRxiv 371500; doi: https://doi.org/10.1101/371500
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A neuron-optimized CRISPR/dCas9 activation system for robust and specific gene regulation
Katherine E. Savell, Svitlana V. Bach, Morgan E. Zipperly, Jasmin S. Revanna, Nicholas A. Goska, Jennifer J. Tuscher, Corey G. Duke, Faraz A. Sultan, Julia N. Burke, Derek M. Williams, Lara Ianov, Jeremy J Day
bioRxiv 371500; doi: https://doi.org/10.1101/371500

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