ErbB2/HER2 CRISPR Activation Plasmid (m): sc-420218-ACT

Mouse Erbb2 encodes the receptor tyrosine kinase ErbB2/HER2, a member of the EGFR/ErbB family that functions as a preferred heterodimerization partner to amplify growth factor signaling. Upon activation, ErbB2 engages MAPK/ERK and PI3K–AKT pathways to regulate proliferation, survival, metabolism, and differentiation, with additional effects on cell adhesion and motility. In mammary epithelium and other tissues, ErbB2 signaling contributes to developmental patterning and tissue homeostasis, and dysregulation is widely used as a model for oncogenic signaling and altered receptor trafficking. Because ErbB2 lacks a known soluble ligand, its activity is often studied through changes in expression levels, dimer composition, and downstream phosphorylation networks.

ErbB2/HER2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Erbb2 expression without altering the underlying DNA sequence.

ErbB2/HER2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Erbb2 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 Erbb2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ErbB2/HER2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Erbb2 locus and enabling the study of ErbB2/HER2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ErbB2/HER2 pathway restoration in tumor cells with silenced or reduced Erbb2 expression.

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