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.
eRF3a CRISPR/Cas9 KO Plasmid (h): sc-404748
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eRF3a 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 eRF3a 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 | |
eRF3a CRISPR/Cas9 KO Plasmid (h) | sc-404748 | 20 µg | $397.00 |
Overview
GSPT1 encodes eukaryotic release factor 3a (eRF3a), a ribosome-associated GTPase that partners with eRF1 to drive translation termination and promote ribosome recycling after stop codon recognition. Through regulated GTP hydrolysis and interactions with the poly(A)-binding protein and other translation factors, eRF3a helps coordinate mRNA surveillance, proteostasis, and the fidelity of protein synthesis. Altered control of translation termination and RNA quality-control pathways has been linked to dysregulated cell growth programs and stress responses, making GSPT1 a useful node for studying how translational control interfaces with signaling and homeostatic networks. In biomedical research, perturbing eRF3a can be leveraged to interrogate mechanisms of stop-codon readthrough, nonsense-mediated decay coupling, and vulnerabilities that emerge from impaired proteome maintenance.
eRF3a CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the GSPT1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the GSPT1 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 GSPT1 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 eRF3a protein expression.
This CRISPR knockout system enables efficient generation of GSPT1-deficient cell models for investigation of eRF3a signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting GSPT1 exon(s) critical for eRF3a 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 GSPT1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by eRF3a CRISPR/Cas9 KO Plasmid (h) and eRF3a CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the GSPT1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by eRF3a HDR Plasmid (h) and eRF3a HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by GSPT1 homology arms to support homology-directed repair at defined GSPT1 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.