patched 2 CRISPR/Cas9 KO Plasmid (h): sc-404866
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- patched 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 patched 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
patched 2 CRISPR/Cas9 KO Plasmid (h) | sc-404866 | 20 µg | $397.00 |
Overview
PTCH2 encodes patched 2, a twelve-pass transmembrane receptor that functions as a key negative regulator of Hedgehog signaling by binding Hedgehog ligands and modulating Smoothened activity. Through control of GLI-dependent transcription, PTCH2 contributes to developmental patterning, cell fate decisions, and tissue homeostasis, and its activity intersects with processes such as proliferation, differentiation, and ciliogenesis. Dysregulated Hedgehog pathway signaling and altered PTCH2 function have been associated with oncogenic signaling contexts and congenital or developmental phenotypes, supporting its relevance to pathway-focused mechanistic studies. PTCH2 is also investigated in relation to receptor trafficking and feedback regulation that tune signal amplitude and duration.
patched 2 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PTCH2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PTCH2 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 PTCH2 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 2 protein expression.
This CRISPR knockout system enables efficient generation of PTCH2-deficient cell models for investigation of patched 2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PTCH2 exon(s) critical for patched 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 PTCH2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by patched 2 CRISPR/Cas9 KO Plasmid (h) and patched 2 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PTCH2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by patched 2 HDR Plasmid (h) and patched 2 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PTCH2 homology arms to support homology-directed repair at defined PTCH2 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.