PROT CRISPR/Cas9 KO Plasmid (h): sc-408452
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PROT 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 PROT 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 | |
PROT CRISPR/Cas9 KO Plasmid (h) | sc-408452 | 20 µg | $397.00 |
Overview
SLC6A7 encodes PROT (proline transporter), a plasma membrane sodium- and chloride-dependent transporter in the SLC6 family that mediates high-affinity uptake of L-proline. By controlling intracellular proline availability, PROT can influence amino acid homeostasis, redox balance, and metabolic coupling to pathways such as glutamate cycling and mitochondrial bioenergetics, particularly in neural tissues where proline handling is tightly regulated. Altered proline transport and metabolism have been linked to neurodevelopmental and neuropsychiatric phenotypes through impacts on neurotransmission and cellular stress responses. SLC6A7 is therefore relevant for studying transporter biology, amino acid–dependent signaling, and mechanisms connecting proline flux to neuronal function.
PROT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SLC6A7 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SLC6A7 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 SLC6A7 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 PROT protein expression.
This CRISPR knockout system enables efficient generation of SLC6A7-deficient cell models for investigation of PROT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SLC6A7 exon(s) critical for PROT 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 SLC6A7 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PROT CRISPR/Cas9 KO Plasmid (h) and PROT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SLC6A7 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PROT HDR Plasmid (h) and PROT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SLC6A7 homology arms to support homology-directed repair at defined SLC6A7 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.