IQGAP1 CRISPR Activation Plasmid (m): sc-424375-ACT

Mouse Iqgap1 encodes IQGAP1, a multifunctional scaffold protein that integrates signals from Rho-family GTPases with the actin and microtubule cytoskeleton to coordinate cell polarity, adhesion, and directed migration. IQGAP1 interfaces with pathways involving RAC1/CDC42, MAPK/ERK signaling, and cadherin–catenin complexes, thereby shaping membrane dynamics and cell–cell junction organization. Through these interactions, it influences processes such as cytokinesis, vesicle trafficking, and receptor signaling output. Dysregulated IQGAP1-dependent scaffolding has been linked to altered epithelial integrity, invasive behavior, and aberrant proliferative signaling in disease-relevant models, supporting its utility in mechanistic studies of tissue remodeling and oncogenic signaling networks.

IQGAP1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Iqgap1 expression without altering the underlying DNA sequence.

IQGAP1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Iqgap1 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 Iqgap1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IQGAP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Iqgap1 locus and enabling the study of IQGAP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IQGAP1 pathway restoration in tumor cells with silenced or reduced Iqgap1 expression.

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