CENP-A CRISPR/Cas9 KO Plasmid (h): sc-402359
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CENP-A 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 CENP-A 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 | |
CENP-A CRISPR/Cas9 KO Plasmid (h) | sc-402359 | 20 µg | $397.00 |
Overview
CENPA encodes CENP-A, a histone H3 variant that replaces canonical H3 at centromeres to establish centromeric chromatin and specify kinetochore identity. CENP-A deposition and maintenance are coordinated with centromere assembly pathways and mitotic cell cycle progression, supporting accurate chromosome alignment and segregation. Perturbation of CENP-A levels or centromere localization destabilizes kinetochore function, promotes chromosomal instability and aneuploidy, and is frequently studied in the context of genome maintenance defects observed across many cancer models.
CENP-A CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CENPA gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CENPA 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 CENPA 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 CENP-A protein expression.
This CRISPR knockout system enables efficient generation of CENPA-deficient cell models for investigation of CENP-A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting CENPA exon(s) critical for CENP-A 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 CENPA genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CENP-A CRISPR/Cas9 KO Plasmid (h) and CENP-A CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CENPA locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CENP-A HDR Plasmid (h) and CENP-A HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CENPA homology arms to support homology-directed repair at defined CENPA 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.