β-casein CRISPR Activation Plasmid (h): sc-402248-ACT

CSN2 encodes human β-casein, a major milk protein synthesized by differentiated mammary epithelial cells and commonly used as a readout of lactogenic differentiation. β-casein expression is regulated by endocrine and cytokine signaling, integrating prolactin–JAK2/STAT5 activation with glucocorticoid and insulin pathways to coordinate transcriptional programs during mammary gland development. In model systems, CSN2 serves as a functional marker of secretory activation and epithelial maturation, linking chromatin state and enhancer usage to lineage-specific gene expression. Dysregulation of these differentiation-associated networks is relevant to studies of mammary epithelial plasticity, stress responses, and altered transcriptional control observed in breast disease biology.

β-casein CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CSN2 expression without altering the underlying DNA sequence.

β-casein CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CSN2 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 CSN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β-casein expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CSN2 locus and enabling the study of β-casein-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β-casein pathway restoration in tumor cells with silenced or reduced CSN2 expression.

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