Work

An Alternative Gene Editing Strategy Using a Single AAV Vector

Year: 2025

Type: article

Abstract: (SpCas9) and Cre recombinase to enable selective co-expression of standard neuroscience tools in edited cells. This protocol can produce AAVs of any serotype, and the resulting AAVs can be used in the central and peripheral nervous systems. This flexible approach could help identify and test the function of novel genes affecting synaptic transmission, circuit activity, or morphology with a single viral injection. Key features • Single-vector CRISPR/Cas9 gene editing and genetically encoded tool delivery for use in mouse central and peripheral nervous systems. • Can be combined with many genetically encoded tools, including fluorescent proteins, optogenetic and chemogenetic tools, and calcium imaging. • Requires first-generation cross between a Cre driver mouse line and Cre-dependent SpCas9 mouse line. • Optimized for use with SpCas9.

Source: BIO-PROTOCOL

DOI: 10.21769/bioprotoc.5362

Authors: Jamie Moffa, Vani Kalyanaraman, Bryan Copits

Institution: Washington University in St. Louis

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: diamond

Funder: National Institute of Neurological Disorders and Stroke

Award: G2493842201