SPPL3 CRISPR/Cas9 KO Plasmid (h): sc-414174
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SPPL3 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 SPPL3 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 | |
SPPL3 CRISPR/Cas9 KO Plasmid (h) | sc-414174 | 20 µg | $397.00 |
Overview
Signal peptide peptidase-like 3 (SPPL3) is an intramembrane aspartyl protease of the GxGD family that cleaves type II membrane proteins within the secretory pathway. It contributes to protein ectodomain shedding and regulates the processing of multiple substrates, influencing Golgi and ER-associated trafficking, glycoprotein maturation, and cell-surface presentation of membrane proteins. SPPL3 activity has been linked to control of N-glycosylation by modulating glycosyltransferase abundance and localization, connecting it to proteostasis and membrane protein quality control. Dysregulation of these processes is relevant to studies of immune signaling, inflammation, and cancer-associated changes in glycosylation and cell-surface receptor function.
SPPL3 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SPPL3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SPPL3 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 SPPL3 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 SPPL3 protein expression.
This CRISPR knockout system enables efficient generation of SPPL3-deficient cell models for investigation of SPPL3 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SPPL3 exon(s) critical for SPPL3 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 SPPL3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by SPPL3 CRISPR/Cas9 KO Plasmid (h) and SPPL3 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SPPL3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by SPPL3 HDR Plasmid (h) and SPPL3 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SPPL3 homology arms to support homology-directed repair at defined SPPL3 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.