Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
TWIK-2 CRISPR/Cas9 KO Plasmid (h): sc-407263
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TWIK-2 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 TWIK-2 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: TWIK-2 Antibody (H-6): sc-376515
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TWIK-2 CRISPR/Cas9 KO Plasmid (h) | sc-407263 | 20 µg | $397.00 |
Overview
KCNK6 encodes TWIK-2 (K2P6.1), a two-pore domain potassium channel that contributes to background (leak) K+ conductance and helps set resting membrane potential and membrane excitability. TWIK-2 activity influences ion homeostasis and calcium-dependent signaling by shaping cellular polarization, with reported roles in vascular smooth muscle function, endothelial biology, and immune cell activation. Through its impact on electrical properties and downstream signaling, TWIK-2 has been studied in pathways linked to vascular tone regulation and inflammatory responses. Dysregulated K2P channel function, including TWIK-2, is therefore relevant to mechanistic investigations of hypertension, vascular dysfunction, and immune-mediated pathophysiology.
TWIK-2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the KCNK6 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the KCNK6 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 KCNK6 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 TWIK-2 protein expression.
This CRISPR knockout system enables efficient generation of KCNK6-deficient cell models for investigation of TWIK-2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting KCNK6 exon(s) critical for TWIK-2 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 KCNK6 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TWIK-2 CRISPR/Cas9 KO Plasmid (h) and TWIK-2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the KCNK6 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TWIK-2 HDR Plasmid (h) and TWIK-2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by KCNK6 homology arms to support homology-directed repair at defined KCNK6 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.