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.
patched/PTCH1 CRISPR/Cas9 KO Plasmid (h): sc-400457
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- patched/PTCH1 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 patched/PTCH1 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: patched/PTCH1 Antibody (A-2): sc-518044
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
patched/PTCH1 CRISPR/Cas9 KO Plasmid (h) | sc-400457 | 20 µg | $397.00 |
Overview
PTCH1 encodes patched, a 12-pass transmembrane receptor that restrains Hedgehog signaling by inhibiting SMO in the absence of ligand. Binding of Sonic/Indian/Desert Hedgehog relieves this repression, enabling GLI-dependent transcriptional programs that govern embryonic patterning, tissue homeostasis, and cell fate decisions. PTCH1 activity links membrane sterol transport and ciliary trafficking to pathway output, integrating cues that modulate proliferation and differentiation. Dysregulated PTCH1-Hedgehog signaling is implicated in developmental disorders and oncogenic processes, making PTCH1 a key node for pathway and genotype–phenotype studies.
patched/PTCH1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PTCH1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PTCH1 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 PTCH1 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 patched/PTCH1 protein expression.
This CRISPR knockout system enables efficient generation of PTCH1-deficient cell models for investigation of patched/PTCH1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PTCH1 exon(s) critical for patched/PTCH1 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 PTCH1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by patched/PTCH1 CRISPR/Cas9 KO Plasmid (h) and patched/PTCH1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PTCH1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by patched/PTCH1 HDR Plasmid (h) and patched/PTCH1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PTCH1 homology arms to support homology-directed repair at defined PTCH1 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.