Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
PCMT1 CRISPR/Cas9 KO Plasmid (h): sc-418153
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PCMT1 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 PCMT1 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
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PCMT1 Antibody (D-E7): sc-100977
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PCMT1 CRISPR/Cas9 KO Plasmid (h) | sc-418153 | 20 µg | $397.00 |
Overview
PCMT1 encodes protein-L-isoaspartate (D-aspartate) O-methyltransferase 1, a SAM-dependent methyltransferase that repairs age- or stress-induced isoaspartyl damage in proteins by methylating abnormal Asp/Asn residues to facilitate conversion back to normal linkages. This protein quality-control activity supports proteostasis and limits accumulation of dysfunctional, aggregation-prone proteins, linking PCMT1 to cellular responses to oxidative stress and long-lived protein maintenance in the cytosol and nucleus. PCMT1 function intersects with pathways governing protein turnover, chaperone networks, and stress resilience, making it relevant to studies of protein aging and post-translational damage. Altered PCMT1 activity has been investigated in contexts of neurobiology and other disorders where protein damage and impaired repair mechanisms contribute to cellular dysfunction.
PCMT1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PCMT1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PCMT1 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 PCMT1 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 PCMT1 protein expression.
This CRISPR knockout system enables efficient generation of PCMT1-deficient cell models for investigation of PCMT1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PCMT1 exon(s) critical for PCMT1 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 PCMT1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PCMT1 CRISPR/Cas9 KO Plasmid (h) and PCMT1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PCMT1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PCMT1 HDR Plasmid (h) and PCMT1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PCMT1 homology arms to support homology-directed repair at defined PCMT1 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.