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.
DNA pol ε B CRISPR/Cas9 KO Plasmid (h): sc-418231
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DNA pol ε B 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 DNA pol ε B 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: DNA pol ε B Antibody (C-9): sc-398582
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DNA pol ε B CRISPR/Cas9 KO Plasmid (h) | sc-418231 | 20 µg | $397.00 |
Overview
POLE2 encodes the DNA polymerase epsilon subunit B, an accessory component of the Pol ε holoenzyme that supports high-fidelity DNA synthesis during S phase and contributes to leading-strand replication. Through coupling with the replisome, POLE2 helps coordinate processive DNA elongation, replication fork stability, and DNA damage tolerance, linking DNA replication to checkpoint signaling and genome maintenance pathways. Altered Pol ε function and replication stress are associated with increased mutational burden and chromosomal instability, making POLE2 a useful target for studying mechanisms that shape genome integrity in proliferating human cells.
DNA pol ε B CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the POLE2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POLE2 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 POLE2 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 DNA pol ε B protein expression.
This CRISPR knockout system enables efficient generation of POLE2-deficient cell models for investigation of DNA pol ε B signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting POLE2 exon(s) critical for DNA pol ε B 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 POLE2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by DNA pol ε B CRISPR/Cas9 KO Plasmid (h) and DNA pol ε B CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the POLE2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by DNA pol ε B HDR Plasmid (h) and DNA pol ε B HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by POLE2 homology arms to support homology-directed repair at defined POLE2 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.