Conditional Single Vector CRISPR/SaCas9 Viruses for Efficient Mutagenesis in the Adult Mouse Nervous System
Year: 2020
Type: article
Abstract: Mice engineered for conditional, cell type-specific gene inactivation have dominated the field of mouse genetics because of the high efficiency of Cre-loxP-mediated recombination. Recent advances in CRISPR/Cas9 technologies have provided alternatives for rapid gene mutagenesis for loss-of-function (LOF) analysis. Whether these strategies can be streamlined for rapid genetic analysis with the efficiencies comparable with those of conventional genetic approaches has yet to be established. We show that a single adeno-associated viral (AAV) vector containing a recombinase-dependent Staphylococcus aureus Cas9 (SaCas9) and a single guide RNA (sgRNA) are as efficient as conventional conditional gene knockout and can be adapted for use in either Cre- or Flp-driver mouse lines. The efficacy of this approach is demonstrated for the analysis of GABAergic, glutamatergic, and monoaminergic neurotransmission. Using this strategy, we reveal insight into the role of GABAergic regulation of midbrain GABA-producing neurons in psychomotor activation.
Source: Cell Reports
Institutions: University of Washington, Northwestern University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $5,200
Awards: Discovering the Role of VTA GABA in Dopamine-Dependent Psychomotor Activation through CRISPR/SaCas9 Gene Knockout, Molecular, anatomic, and functional characterization of midbrain dopamine neuron subtypes, Training in Cellular and Molecular Biology, Psychology Training in Alcohol Research (PTAR), Isolation of brain reward circuits using peptidergic systems, Phasic dopamine and symptom domains of mental illness, University of Washington Center of Excellence in Opioid Addiction Research
OpenAlex ID: W3013220673