Ral BP-1 CRISPR Activation Plasmid (h): sc-402752-ACT

RALBP1 encodes Ral BP-1 (also known as RLIP76), a multifunctional effector of Ral GTPases that integrates signaling with membrane trafficking and cytoskeletal dynamics. Ral BP-1 participates in clathrin-dependent endocytosis, receptor internalization, and actin remodeling through interactions with adaptors and small GTPase pathways, linking RalA/RalB activity to motility and stress-responsive cellular programs. The protein has also been connected to ATP-dependent transport functions and cellular detoxification responses, supporting survival under oxidative or xenobiotic stress. Dysregulated RALBP1 expression or signaling is associated with altered proliferation, invasion, and therapy resistance phenotypes in multiple tumor contexts, making it relevant for mechanistic studies of oncogenic signaling and cellular stress adaptation.

Ral BP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RALBP1 expression without altering the underlying DNA sequence.

Ral BP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RALBP1 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 RALBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ral BP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RALBP1 locus and enabling the study of Ral BP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ral BP-1 pathway restoration in tumor cells with silenced or reduced RALBP1 expression.

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