phapi2 CRISPR/Cas9 KO Plasmid (m): sc-422189
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- phapi2 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 phapi2 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 | |
phapi2 CRISPR/Cas9 KO Plasmid (m) | sc-422189 | 20 µg | $397.00 |
Overview
Mouse Pemt encodes phosphatidylethanolamine N-methyltransferase, a key enzyme in phosphatidylcholine biosynthesis that catalyzes sequential methylation of phosphatidylethanolamine using S-adenosylmethionine. This activity supports membrane phospholipid homeostasis, lipoprotein assembly, and methyl-donor metabolism, linking PEMT to lipid remodeling and cellular stress responses. Pemt-dependent phosphatidylcholine production is particularly relevant to hepatic lipid handling, ER function, and signaling processes that depend on membrane composition. Perturbation of PEMT-mediated pathways has been associated with altered lipid metabolism phenotypes and provides a tractable model for studying metabolic and membrane-associated mechanisms.
phapi2 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Pemt gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Pemt 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 Pemt 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 phapi2 protein expression.
This CRISPR knockout system enables efficient generation of Pemt-deficient cell models for investigation of phapi2 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Pemt exon(s) critical for phapi2 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 Pemt genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by phapi2 CRISPR/Cas9 KO Plasmid (m) and phapi2 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Pemt locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by phapi2 HDR Plasmid (m) and phapi2 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Pemt homology arms to support homology-directed repair at defined Pemt 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.