eIF3δ CRISPR/Cas9 KO Plasmid (h): sc-407744
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eIF3δ 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 eIF3δ genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- eIF3δ HDR Plasmid (h) (sc-407744-HDR) is recommended for co-transfection with eIF3δ CRISPR/Cas9 KO Plasmid (h) to enable selection of successfully edited cells through HDR-mediated integration of a puromycin resistance cassette and RFP reporter gene
- eIF3δ HDR Plasmid (h) is a pool of plasmids, each containing a homology-directed repair (HDR) template corresponding to the gRNA target sites in the eIF3δ CRISPR/Cas9 KO Plasmid (h)
- Each HDR plasmid contains two ~800 bp homology arms flanking the puromycin resistance and RFP cassettes, designed to bind genomic DNA sequences surrounding the Cas9-induced double-strand break site and facilitate precise HDR-mediated integration
- 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 | |
eIF3δ CRISPR/Cas9 KO Plasmid (h) | sc-407744 | 20 µg | $397.00 | |||
eIF3δ HDR Plasmid (h) | sc-407744-HDR | 20 µg | $445.00 |
Overview
EIF3G encodes the human eIF3δ subunit of the eukaryotic translation initiation factor 3 (eIF3) complex, a central scaffold that coordinates 40S ribosome recruitment and translation initiation on mRNAs. Through interactions with initiation factors and the 43S preinitiation complex, eIF3δ contributes to global protein synthesis as well as transcript-selective translation programs that shape proliferation, stress adaptation, and cell-cycle control. Altered translational control involving eIF3 components has been linked to dysregulated growth signaling and oncogenic phenotypes, making EIF3G a useful node for studying how translation initiation interfaces with pathways such as mTOR-dependent protein synthesis and integrated stress responses. EIF3G perturbation is therefore relevant for mechanistic studies of proteostasis, ribosome loading dynamics, and disease-associated translational rewiring in human cells.
eIF3δ CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the EIF3G gene in human cell lines. Each plasmid in the pool co-expresses a unique sgRNA, targeting a distinct site within the EIF3G locus, alongside the Streptococcus pyogenes Cas9 nuclease, and encodes GFP to enable fluorescent identification and enrichment of successfully transfected cells. This multi-guide strategy increases the likelihood of inducing frameshifts or deletions that produce a functional knockout, offering a more robust alternative to single-guide approaches. DSBs induced at multiple sites are resolved through non-homologous end joining (NHEJ) or, when used with the included HDR donor template, homology-directed repair (HDR) at a defined target site within the locus.
When used in conjunction with the RFP-expressing HDR donor, GFP and RFP fluorescence can be used together to distinguish transfected from edited cell populations, streamlining flow cytometry-based sorting and clone selection workflows.
Homology-Directed Repair (HDR) Donor — Puromycin Cassette with RFP Reporter
For applications requiring confirmed, selectable knockout clones, eIF3δ HDR Plasmid (h) includes an HDR donor construct containing a puromycin resistance cassette (PuroR) and a red fluorescent protein (RFP) reporter, flanked by homology arms specific to a defined EIF3G target site.
When co-transfected with eIF3δ CRISPR/Cas9 KO Plasmid (h):
- The PuroR-RFP cassette integrates at the Cas9 cut site via HDR, disrupting the EIF3G open reading frame.
- RFP fluorescence provides an immediate visual indicator of successful integration, enabling fluorescence-based identification or sorting of edited cells prior to or alongside puromycin selection.
- Successfully edited cells are confirmed through puromycin resistance, substantially reducing clone screening burden.
- This selection strategy is ideal for generating stable, clonal KO cell lines for downstream functional studies, drug screening, or model development.
Cre-lox Cassette Removal System
The HDR donor construct features loxP sites flanking the PuroR-RFP selection cassette to allow clean marker removal following clone confirmation. Transient expression of Cre recombinase via the included Cre Vector: sc-418923 excises the cassette, leaving a minimal residual loxP site within the EIF3G locus and eliminating potential confounding effects on downstream assays.
This two-step approach:
- Minimizes disruption to local chromatin architecture and neighboring regulatory elements
- Restores a near-native genomic context at the edited locus
- Enables reuse of the puromycin selection strategy in the same cell line for additional edits
Key Features
- gRNA targeting EIF3G exon(s) critical for eIF3δ function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- HDR donor with puromycin resistance for positive clone selection
- loxP-flanked PuroR cassette with Cre recombinase vector for seamless marker removal
- Supplied ready to use for delivery by transfection
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.