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eNeuro. 2019 Mar 7;6(1). pii: ENEURO.0495-18.2019. doi: 10.1523/ENEURO.0495-18.2019. eCollection 2019 Jan-Feb.

A Neuron-Optimized CRISPR/dCas9 Activation System for Robust and Specific Gene Regulation.

Author information

1
Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of Alabama at Birmingham, Birmingham, AL, 35294.
2
Civitan International Research Center, University of Alabama at Birmingham, Birmingham, AL, 35294.

Abstract

CRISPR-based technology has provided new avenues to interrogate gene function, but difficulties in transgene expression in post-mitotic neurons has delayed incorporation of these tools in the central nervous system (CNS). Here, we demonstrate a highly efficient, neuron-optimized dual lentiviral CRISPR-based transcriptional activation (CRISPRa) system capable of robust, modular, and tunable gene induction and multiplexed gene regulation across several primary rodent neuron culture systems. 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.

KEYWORDS:

Bdnf; CRISPR; epigenetics; gene expression; transcription

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