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Abstract
A protocol for constructing a vector expressing three guide RNAs targeting a single gene in the mouse brain is outlined.
- The protocol facilitates the design of CRISPR knockout guide RNAs that target one specific gene.
- Steps include plasmid construction and verification for effective gene editing.
- Neonatal adeno-associated viral (AAV) vectors are utilized for delivery into the mouse brain.
- Brain preparation and immunofluorescence are employed to validate gene disruption.
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