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.
FEN-1 CRISPR/Cas9 KO Plasmid (h): sc-403168
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FEN-1 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 FEN-1 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: FEN-1 Antibody (B-4): sc-28355
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FEN-1 CRISPR/Cas9 KO Plasmid (h) | sc-403168 | 20 µg | $397.00 |
Overview
FEN1 encodes flap endonuclease 1 (FEN-1), a structure-specific nuclease that cleaves 5′ flap intermediates during Okazaki fragment maturation and long-patch base excision repair. FEN-1 coordinates with PCNA and DNA polymerases to support DNA replication fidelity, processing of stalled replication forks, and resolution of secondary DNA structures that can trigger genome instability. Through its roles in replication stress responses, repair pathway choice, and maintenance of chromosomal integrity, altered FEN1 function is linked to mutational accumulation and signatures associated with cancer biology and DNA damage sensitivity. FEN-1 is therefore widely used as a mechanistic entry point for studying DNA repair networks, replication-coupled processing, and checkpoint signaling.
FEN-1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the FEN1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the FEN1 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 FEN1 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 FEN-1 protein expression.
This CRISPR knockout system enables efficient generation of FEN1-deficient cell models for investigation of FEN-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting FEN1 exon(s) critical for FEN-1 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 FEN1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by FEN-1 CRISPR/Cas9 KO Plasmid (h) and FEN-1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the FEN1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by FEN-1 HDR Plasmid (h) and FEN-1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by FEN1 homology arms to support homology-directed repair at defined FEN1 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.