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.
EFP CRISPR/Cas9 KO Plasmid (h): sc-402678
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EFP 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 EFP 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: EFP Antibody (E-4): sc-166926
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EFP CRISPR/Cas9 KO Plasmid (h) | sc-402678 | 20 µg | $397.00 |
Overview
TRIM25 encodes the E3 ubiquitin ligase EFP, a TRIM family protein that catalyzes ubiquitination events controlling innate immune signaling and RNA metabolism. EFP promotes antiviral responses by ubiquitinating and activating RIG-I, facilitating downstream type I interferon signaling through MAVS, TBK1, and IRF3/7, and it can modulate NF-κB-associated inflammatory outputs. Beyond host defense, TRIM25 participates in post-transcriptional regulation via interactions with RNA-binding proteins and influences cell-cycle and stress-response programs. Dysregulated TRIM25 activity has been linked to altered interferon tone, viral susceptibility, and context-dependent roles in tumor biology and endocrine signaling, making it a relevant node in inflammation and cancer research.
EFP CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TRIM25 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TRIM25 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 TRIM25 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 EFP protein expression.
This CRISPR knockout system enables efficient generation of TRIM25-deficient cell models for investigation of EFP signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TRIM25 exon(s) critical for EFP 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 TRIM25 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by EFP CRISPR/Cas9 KO Plasmid (h) and EFP CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TRIM25 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by EFP HDR Plasmid (h) and EFP HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TRIM25 homology arms to support homology-directed repair at defined TRIM25 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.