GlyT2 CRISPR/Cas9 KO Plasmid (h): sc-403077
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GlyT2 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 GlyT2 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: GlyT2 Antibody (B-4): sc-390090
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GlyT2 CRISPR/Cas9 KO Plasmid (h) | sc-403077 | 20 µg | $397.00 |
Overview
SLC6A5 encodes the presynaptic glycine transporter GlyT2 (SLC6 family), a Na\+/Cl\--dependent transporter that mediates high-affinity reuptake of glycine into inhibitory glycinergic neurons. By replenishing cytosolic glycine for vesicular loading, GlyT2 supports fast synaptic inhibition in the spinal cord and brainstem and helps shape network excitability. This transporter interfaces with neurotransmitter cycling and chloride homeostasis processes that regulate inhibitory tone and sensorimotor circuit output. Disruption or dysregulation of SLC6A5 has been linked to impaired glycinergic signaling and neurophysiological phenotypes relevant to hereditary hyperekplexia and related inhibitory synapse dysfunction.
GlyT2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SLC6A5 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SLC6A5 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 SLC6A5 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 GlyT2 protein expression.
This CRISPR knockout system enables efficient generation of SLC6A5-deficient cell models for investigation of GlyT2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SLC6A5 exon(s) critical for GlyT2 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 SLC6A5 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by GlyT2 CRISPR/Cas9 KO Plasmid (h) and GlyT2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SLC6A5 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by GlyT2 HDR Plasmid (h) and GlyT2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SLC6A5 homology arms to support homology-directed repair at defined SLC6A5 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.