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.
4E-BP1 CRISPR/Cas9 KO Plasmid (m): sc-420149
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 4E-BP1 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 4E-BP1 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: 4E-BP1 Antibody (P-1): sc-9977
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
4E-BP1 CRISPR/Cas9 KO Plasmid (m) | sc-420149 | 20 µg | $397.00 |
Overview
Mouse Eif4ebp1 encodes 4E-BP1, a cap-dependent translation repressor that binds EIF4E and limits eIF4F complex assembly, thereby tuning mRNA translation initiation in response to nutrient and growth factor cues. 4E-BP1 is a core downstream effector of the PI3K–AKT–mTORC1 axis; phosphorylation by mTORC1 relieves EIF4E sequestration and promotes selective translation of transcripts involved in cell growth, metabolism, and stress adaptation. Through this translational checkpoint, Eif4ebp1 contributes to regulation of autophagy, cell cycle progression, and proteostasis programs. Dysregulated 4E-BP1 signaling is widely used as a molecular readout of aberrant mTOR pathway activity in models of cancer biology, metabolic dysfunction, and neurodevelopmental and neurodegenerative processes.
4E-BP1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Eif4ebp1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Eif4ebp1 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 Eif4ebp1 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 4E-BP1 protein expression.
This CRISPR knockout system enables efficient generation of Eif4ebp1-deficient cell models for investigation of 4E-BP1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Eif4ebp1 exon(s) critical for 4E-BP1 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 Eif4ebp1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by 4E-BP1 CRISPR/Cas9 KO Plasmid (m) and 4E-BP1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Eif4ebp1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by 4E-BP1 HDR Plasmid (m) and 4E-BP1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Eif4ebp1 homology arms to support homology-directed repair at defined Eif4ebp1 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.