eIF1B CRISPR Activation Plasmid (h): sc-403809-ACT

EIF1B encodes the human eukaryotic translation initiation factor 1B (eIF1B), a conserved component of the translation initiation machinery that supports start-codon selection and efficient assembly of initiation complexes. By influencing global protein synthesis capacity, eIF1B intersects with cellular programs that depend on precise translational control, including cell-cycle progression, stress responses, and adaptation to nutrient and growth-factor signaling. Dysregulated translation initiation is a recurrent feature of proliferative and stress-adaptive states, making EIF1B relevant for mechanistic studies of gene-expression regulation in disease-associated contexts. EIF1B perturbation can therefore be used to probe how altered initiation fidelity and translational output reshape downstream pathways and phenotypes.

eIF1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EIF1B expression without altering the underlying DNA sequence.

eIF1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EIF1B 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 EIF1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous eIF1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EIF1B locus and enabling the study of eIF1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of eIF1B pathway restoration in tumor cells with silenced or reduced EIF1B expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.