κ-casein CRISPR/Cas9 KO Plasmid (h): sc-406821
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- κ-casein 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 κ-casein 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 | |
κ-casein CRISPR/Cas9 KO Plasmid (h) | sc-406821 | 20 µg | $397.00 |
Overview
CSN3 encodes κ-casein, a secreted phosphoprotein classically characterized as a component of milk casein micelles where it contributes to protein solubility and calcium phosphate organization. As an extracellular glycoprotein, κ-casein undergoes post-translational modifications that influence protein–protein interactions and secretion dynamics, providing a model substrate for studying secretory pathway processing and proteolytic cleavage. Although CSN3 biology is best defined in lactation-related contexts, variation in κ-casein sequence and glycosylation has been associated with altered protein stability and bioactive peptide generation, making it relevant for mechanistic studies of extracellular protein processing and signaling. In human systems, CSN3 serves as a tractable locus for interrogating regulation of secreted protein expression and the downstream effects of proteolysis-derived peptides on cellular responses.
κ-casein CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CSN3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CSN3 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 CSN3 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 κ-casein protein expression.
This CRISPR knockout system enables efficient generation of CSN3-deficient cell models for investigation of κ-casein signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CSN3 exon(s) critical for κ-casein 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 CSN3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by κ-casein CRISPR/Cas9 KO Plasmid (h) and κ-casein CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CSN3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by κ-casein HDR Plasmid (h) and κ-casein HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CSN3 homology arms to support homology-directed repair at defined CSN3 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.