Elastase-1 CRISPR Activation Plasmid (h): sc-402707-ACT

Human CELA1 encodes elastase-1, a secreted serine protease that catalyzes proteolytic remodeling of extracellular matrix components and contributes to regulated peptide processing in mucosal and glandular tissues. Elastase-1 activity intersects with protease–antiprotease balance, inflammatory signaling, and tissue remodeling processes that influence epithelial integrity and stromal interactions. Dysregulated elastase activity has been associated with pathological matrix degradation and chronic inflammatory microenvironments, making CELA1 relevant to studies of airway remodeling, fibrosis-associated proteolysis, and tumor–stroma crosstalk. As a digestive- and barrier-adjacent protease, CELA1 is also useful for interrogating secretory pathway dynamics and protease-driven signaling networks.

Elastase-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CELA1 expression without altering the underlying DNA sequence.

Elastase-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CELA1 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 CELA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Elastase-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CELA1 locus and enabling the study of Elastase-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Elastase-1 pathway restoration in tumor cells with silenced or reduced CELA1 expression.

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