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

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.

SUBMITTER: Savell KE 

PROVIDER: S-EPMC6412672 | biostudies-literature | 2019 Jan-Feb

REPOSITORIES: biostudies-literature