U6-(gRNA)-hSyn-Cre-P2A-EGFP-KASH-hGHpA
Single-gRNA vector expressing Cre recombinase and EGFP-KASH from the neuron-specific hSyn promoter via P2A self-cleavage, enabling simultaneous Cre/loxP recombination and FANS-compatible nuclear envelope labeling of transduced neurons. Adapted from Addgene #60231 (Platt et al. 2014, Feng Zhang lab) with hGHpA substituted for the original shortPA.
Characteristics
Dual-function single-AAV design. hSyn-driven Cre-P2A-EGFP-KASH produces both functional Cre recombinase and perinuclear EGFP signal from a single ORF via P2A cleavage. EGFP-KASH localizes to the outer nuclear membrane enabling FANS-based nuclei isolation. Neuronal specificity is conferred by hSyn — non-neuronal cells receive the gRNA but express neither Cre nor the reporter.
Custom Inserts
Literature Sources
- Platt et al. (2014). CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell- Platt 2014 Cas9 Mice
Available Products
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Additional Information
Validated in: Mouse neurons in vivo (cortex, hippocampus); Cas9 knock-in mouse lines
Applications: In vivo neuronal CRISPR editing in Cas9 knock-in mice (Rosa26-Cas9 or equivalent) where Cre-dependent recombination of a loxP reporter or conditional allele is needed alongside gRNA delivery. EGFP-KASH enables FANS-based sorting of transduced neuronal nuclei for downstream snRNA-seq or snATAC-seq.