4E-BP1 CRISPR Activation Plasmid (h): sc-400398-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 4E-BP1 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- 4E-BP1 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by 4E-BP1 CRISPR Activation Plasmid (h) and 4E-BP1 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the EIF4EBP1 transcriptional start site. One or both designs may be available
- 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 Activation Plasmid (h) | sc-400398-ACT | 20 µg | $397.00 | |||
4E-BP1 CRISPR Activation Plasmid (h2) | sc-400398-ACT-2 | 20 µg | $397.00 |
EIF4EBP1 encodes 4E-BP1, a phosphorylation-sensitive repressor of cap-dependent translation that binds EIF4E and limits eIF4F complex assembly. As a key downstream effector of PI3K–AKT–mTOR signaling, 4E-BP1 integrates growth factor, nutrient, and stress inputs to tune global protein synthesis, cell growth, and metabolic adaptation. mTORC1-mediated phosphorylation of 4E-BP1 releases EIF4E to promote translation of transcripts involved in proliferation and survival, linking EIF4EBP1 regulation to proteostasis and cell-cycle control. Dysregulated 4E-BP1 activity and phosphorylation state are frequently studied in cancer biology, metabolic signaling, and stress-response contexts where translational rewiring contributes to disease-associated phenotypes.
4E-BP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF4EBP1 expression without altering the underlying DNA sequence.
4E-BP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF4EBP1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the EIF4EBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous 4E-BP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF4EBP1 locus and enabling the study of 4E-BP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of 4E-BP1 pathway restoration in tumor cells with silenced or reduced EIF4EBP1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.