β-defensin 124 CRISPR Activation Plasmid (h): sc-410392-ACT

DEFB124 encodes human β-defensin 124, a small cationic peptide within the defensin family that contributes to epithelial innate immunity through direct antimicrobial activity and modulation of local inflammatory responses. β-defensins participate in mucosal barrier processes and can influence chemotactic signaling and cytokine networks that shape leukocyte recruitment and host–microbe interactions. Altered defensin expression patterns have been associated with dysregulated mucosal immunity and chronic inflammatory states, supporting investigation of DEFB124 in infection biology and immune homeostasis. In reproductive and urogenital tract contexts, defensin family members are also studied for their roles in maintaining barrier integrity and microbiome balance.

β-defensin 124 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DEFB124 expression without altering the underlying DNA sequence.

β-defensin 124 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DEFB124 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 DEFB124 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous β-defensin 124 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DEFB124 locus and enabling the study of β-defensin 124-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of β-defensin 124 pathway restoration in tumor cells with silenced or reduced DEFB124 expression.

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