EAAT4 CRISPR/Cas9 KO Plasmid (m): sc-422986
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EAAT4 CRISPR/Cas9 Knockout (KO) Plasmid (m) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the EAAT4 genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EAAT4 Antibody (6D9): sc-293344
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EAAT4 CRISPR/Cas9 KO Plasmid (m) | sc-422986 | 20 µg | $397.00 |
Overview
Slc1a6 encodes excitatory amino acid transporter 4 (EAAT4), a high-affinity Na+-dependent glutamate transporter that contributes to extracellular glutamate clearance and limits excitotoxic signaling at synapses. In the mouse CNS, EAAT4 is enriched in cerebellar Purkinje neurons and shapes synaptic transmission by coupling glutamate uptake to ionic gradients, thereby influencing neuronal excitability and plasticity. Through regulation of glutamatergic neurotransmission, EAAT4 interfaces with pathways linked to synaptic homeostasis, calcium-dependent signaling, and neuroinflammatory responses to dysregulated glutamate. Altered glutamate transport capacity is relevant to models of cerebellar dysfunction and broader neurodegeneration-associated phenotypes where excitatory signaling and synaptic integrity are perturbed.
EAAT4 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Slc1a6 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Slc1a6 together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the Slc1a6 open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish EAAT4 protein expression.
This CRISPR knockout system enables efficient generation of Slc1a6-deficient cell models for investigation of EAAT4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Slc1a6 exon(s) critical for EAAT4 function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple Slc1a6 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by EAAT4 CRISPR/Cas9 KO Plasmid (m) and EAAT4 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Slc1a6 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by EAAT4 HDR Plasmid (m) and EAAT4 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Slc1a6 homology arms to support homology-directed repair at defined Slc1a6 target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.