CAMP CRISPR Activation Plasmid (h): sc-400651-ACT

Human CAMP encodes cathelicidin antimicrobial peptide (LL-37), an innate immune effector produced by epithelial cells and myeloid lineages that provides direct antimicrobial activity and shapes mucosal barrier defense. Beyond microbicidal functions, CAMP modulates inflammatory signaling by influencing chemotaxis, cytokine production, and neutrophil and monocyte responses, with connections to pattern-recognition receptor pathways and tissue repair programs. Dysregulated CAMP expression has been associated with inflammatory skin and airway conditions, altered host–microbiome interactions, and susceptibility to infection, making it a useful node for studying immune homeostasis and epithelial–immune crosstalk.

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

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

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