BTC CRISPR Activation Plasmid (h): sc-401927-ACT

Human BTC encodes betacellulin, an epidermal growth factor family ligand that is synthesized as a membrane-anchored precursor and released by proteolytic shedding to signal in autocrine and paracrine contexts. BTC activates EGFR/ERBB receptors to engage downstream MAPK/ERK and PI3K/AKT signaling, shaping cell proliferation, survival, migration, and epithelial–mesenchymal interactions. Its expression is regulated during tissue remodeling and inflammatory signaling and has been associated with altered growth-factor circuitry in cancer biology and cardiometabolic and endocrine-relevant contexts, including pancreatic islet function. As a secreted/membrane-tethered growth factor, BTC is frequently used to study receptor–ligand dynamics, transcriptional programs, and pathway crosstalk that influence cell-state transitions.

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

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

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