KCC2 CRISPR/Cas9 KO Plasmid (h): sc-402080
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- KCC2 CRISPR/Cas9 Knockout (KO) Plasmid (h) 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 KCC2 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
KCC2 CRISPR/Cas9 KO Plasmid (h) | sc-402080 | 20 µg | $397.00 |
Overview
SLC12A5 encodes the neuron-specific K+-Cl− cotransporter KCC2, a key regulator of intracellular chloride homeostasis that enables hyperpolarizing GABAergic and glycinergic neurotransmission. By driving Cl− extrusion, KCC2 shapes inhibitory synaptic strength, membrane excitability, and the maturation of neural circuits, and it interfaces functionally with ion transport and synaptic signaling pathways that govern activity-dependent plasticity. Altered KCC2 expression or function has been associated with disrupted excitation–inhibition balance in the central nervous system, supporting relevance to studies of epilepsy, neurodevelopmental disorders, neuropathic pain, and neurodegeneration. KCC2 is also used as a marker and mechanistic node for investigating inhibitory synapse development, chloride dynamics, and network oscillations.
KCC2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SLC12A5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SLC12A5 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 SLC12A5 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 KCC2 protein expression.
This CRISPR knockout system enables efficient generation of SLC12A5-deficient cell models for investigation of KCC2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SLC12A5 exon(s) critical for KCC2 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 SLC12A5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by KCC2 CRISPR/Cas9 KO Plasmid (h) and KCC2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SLC12A5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by KCC2 HDR Plasmid (h) and KCC2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SLC12A5 homology arms to support homology-directed repair at defined SLC12A5 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.