PLUNC CRISPR Activation Plasmid (h): sc-416774-ACT

BPIFA1 encodes PLUNC (palate, lung, and nasal epithelium clone), a secreted innate immune protein enriched in upper airway epithelium and airway surface liquid. PLUNC contributes to mucosal host defense by modulating antimicrobial activity, influencing biofilm formation, and regulating airway surface tension and epithelial barrier homeostasis. BPIFA1 expression is linked to epithelial differentiation programs and inflammatory signaling in the respiratory tract, making it relevant to studies of chronic airway inflammation and infection biology. Altered PLUNC levels have been associated with airway diseases characterized by dysregulated mucosal immunity, including asthma, chronic obstructive pulmonary disease, and chronic rhinosinusitis.

PLUNC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BPIFA1 expression without altering the underlying DNA sequence.

PLUNC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BPIFA1 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 BPIFA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLUNC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BPIFA1 locus and enabling the study of PLUNC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLUNC pathway restoration in tumor cells with silenced or reduced BPIFA1 expression.

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