PAM CRISPR/Cas9 KO Plasmid (m): sc-422110
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PAM CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 PAM 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 | |
PAM CRISPR/Cas9 KO Plasmid (m) | sc-422110 | 20 µg | $397.00 |
Overview
Peptidylglycine alpha-amidating monooxygenase (PAM), encoded by the mouse Pam gene, is a bifunctional enzyme that catalyzes C-terminal amidation of many neuropeptides and peptide hormones, a modification required for full bioactivity. This copper- and ascorbate-dependent pathway supports peptide maturation within the secretory pathway and influences regulated secretion in neuroendocrine and neuronal cells. Pam activity links vesicle biogenesis and secretory granule function to neuropeptide signaling networks that modulate synaptic transmission, stress responses, metabolism, and cardiovascular homeostasis. Dysregulated peptide amidation and altered PAM function have been associated with neurodevelopmental and neuropsychiatric phenotypes, endocrine imbalance, and cardiometabolic traits, making Pam a useful locus for mechanistic studies of secretory pathway biology.
PAM CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pam gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pam 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 Pam 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 PAM protein expression.
This CRISPR knockout system enables efficient generation of Pam-deficient cell models for investigation of PAM signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pam exon(s) critical for PAM 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 Pam genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PAM CRISPR/Cas9 KO Plasmid (m) and PAM CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pam locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PAM HDR Plasmid (m) and PAM HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pam homology arms to support homology-directed repair at defined Pam 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.