ErbB4/HER4 CRISPR Activation Plasmid (h): sc-400275-ACT

ERBB4 encodes the receptor tyrosine kinase ErbB4/HER4, a member of the EGFR/ErbB family that transduces extracellular cues from neuregulins and related ligands. Upon activation, ErbB4 participates in receptor dimerization and autophosphorylation, propagating signaling through MAPK/ERK, PI3K/AKT, JAK/STAT, and PLCγ pathways to regulate proliferation, differentiation, migration, and survival. ERBB4 is broadly studied in developmental and tissue homeostasis contexts, including neural and epithelial signaling programs, and its dysregulation has been associated with oncogenic signaling networks and altered cell fate control. These pathway connections make ERBB4 a useful node for probing receptor cross-talk, feedback regulation, and context-dependent signaling outputs in human cells.

ErbB4/HER4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ERBB4 expression without altering the underlying DNA sequence.

ErbB4/HER4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ERBB4 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 ERBB4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ErbB4/HER4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ERBB4 locus and enabling the study of ErbB4/HER4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ErbB4/HER4 pathway restoration in tumor cells with silenced or reduced ERBB4 expression.

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