IRSp53 CRISPR Activation Plasmid (h): sc-403072-ACT

BAIAP2 encodes IRSp53, an adaptor protein that links membrane curvature sensing to actin cytoskeleton remodeling through interactions with Rho family GTPases such as Cdc42, SH3-domain binding partners, and actin regulatory complexes. IRSp53 contributes to formation of filopodia and lamellipodia, coordinating processes including cell migration, adhesion dynamics, and neurite outgrowth. Through its role in cytoskeletal organization and membrane trafficking, BAIAP2 activity intersects with signaling networks that regulate epithelial and neuronal morphology. Altered IRSp53-dependent actin dynamics has been associated with neurodevelopmental phenotypes and tumor-associated changes in cell motility in mechanistic studies.

IRSp53 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BAIAP2 expression without altering the underlying DNA sequence.

IRSp53 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BAIAP2 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 BAIAP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IRSp53 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BAIAP2 locus and enabling the study of IRSp53-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IRSp53 pathway restoration in tumor cells with silenced or reduced BAIAP2 expression.

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