CIB CRISPR Activation Plasmid (h): sc-405501-ACT

Human CIB1 (calcium and integrin-binding protein 1) encodes CIB, a myristoylated EF-hand calcium-binding regulator that couples Ca²⁺ signals to integrin-dependent adhesion and cytoskeletal remodeling. CIB1 participates in signaling networks that influence focal adhesion dynamics, platelet and endothelial cell responses, and stress-adaptive pathways linked to cell survival and migration. Through interactions with integrin cytoplasmic tails and other signaling proteins, CIB1 can modulate kinase activity and downstream transcriptional programs that affect proliferation and motility. Dysregulated CIB1 expression or signaling has been reported in contexts relevant to vascular biology, thrombosis-related mechanisms, and cancer-associated invasion and metastasis, supporting its use as a mechanistic target in pathway studies.

CIB CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CIB1 expression without altering the underlying DNA sequence.

CIB CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CIB1 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 CIB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CIB expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CIB1 locus and enabling the study of CIB-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CIB pathway restoration in tumor cells with silenced or reduced CIB1 expression.

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