U6-(SaCas9gRNA)-EFS-EGFP-KASH-hGHpA
AAV transfer plasmid that delivers a SaCas9 guide RNA expressed from U6 alongside an EGFP-KASH fusion reporter expressed from EFS. EGFP-KASH localizes to the outer nuclear membrane enabling fluorescence-activated nuclear sorting (FANS) of transduced nuclei. Adapted from Addgene #113694 as the companion gRNA vector to a separately delivered SaCas9 AAV.
Characteristics
Dual-cassette single-AAV design. U6-driven SaCas9 gRNA cassette is constitutively active. EFS-driven EGFP-KASH is expressed independently and localizes to the outer nuclear membrane (perinuclear pattern confirmed by ICC in 293FT cells). GFP signal enables FANS isolation of transduced nuclei for downstream genomic or epigenomic analysis. Titered and used in vivo at 5.0×10¹² GC/mL.
Custom Inserts
Related Designs
Literature Sources
- 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
Validated in: 293FT cells; mouse basal lateral amygdala neurons (in vivo, AAV-DJ serotype)
Applications: Co-infused with a SaCas9-expressing AAV for two-vector in vivo CRISPR editing. Validated for stereotaxic delivery to mouse basal lateral amygdala (BLA) neurons. GFP-KASH reporter enables FACS/FANS-based verification of transduction and sorting of transduced nuclei without dissociating cells. Compatible with constitutive SaCas9 system; Dox-inducible and Cre-dependent gRNA variants exist in same plasmid series (Addgene #113700–113703).