(pro)-SpCas9-SV40pA + 3*{U6-(gRNA)}-hSyn-mCherry-KASH-bGHpA

Triple-gRNA SpCas9 companion vector with neuron-specific hSyn-driven mCherry-KASH for FANS-compatible nuclear sorting in neuronal tissue.

Characteristics

Expresses three gRNAs from tandem U6 promoters. hSyn promoter restricts mCherry-KASH expression to neurons, enabling FANS-based isolation of transduced neuronal nuclei from mixed-cell tissue. Red channel avoids interference with GFP-based imaging or co-expressed green reporters.

Parts

SpCas9 Nuclease mCherry KASH Domain hSyn Promoter bGH PolyA Signal SV40 Late PolyA Signal U6 Promoter

Custom Inserts

SpCas9 gRNA target (20-23 bp) SpCas9 gRNA target (20-23 bp) SpCas9 gRNA target (20-23 bp)

Related Designs

U6-(SaCas9gRNA)-EFS-EGFP-KASH-hGHpA (pro)-SpCas9-SV40pA + 3*{U6-(gRNA)}-CAG-mCherry-bGHpA (pro)-SpCas9-SV40pA + 2*{U6-(gRNA)}-CAG-mCherry-bGHpA (pro)-SpCas9-SV40pA + U6-(gRNA)-CAG-mCherry-bGHpA (pro)-SpCas9-SV40pA + 3*{U6-(gRNA)}-CAG-EGFP-KASH-bGHpA

Literature Sources

  1. Kim et al. (2021). Multiplexed CRISPR-Cas9 system in a single adeno-associated virus to simultaneously knock out redundant clock genes. Sci Rep- Kim 2021 Multiplex AAV
  2. Kumar et al. (2018). The Development of an AAV-Based CRISPR SaCas9 Genome Editing System That Can Be Delivered to Neurons in vivo and Regulated via Doxycycline and Cre-Recombinase. Front Mol Neurosci- Kumar 2018 In Vivo CRISPR

Available Products

No products currently available with this design. Contact us for custom orders.

Additional Information

Applications: Multiplex 3-target neuronal CRISPR editing in vivo where cell-type specificity of the reporter is required. Preferred for brain tissue containing mixed neuronal and non-neuronal populations — only neurons will express mCherry-KASH, ensuring sorted nuclei are neuron-derived. Compatible with snRNA-seq or snATAC-seq after FANS.

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