Loricrin CRISPR Activation Plasmid (h): sc-402452-ACT

Human LOR encodes loricrin, a highly abundant structural protein of the cornified envelope that supports epidermal barrier formation during terminal keratinocyte differentiation. Loricrin becomes extensively crosslinked by transglutaminases and integrates with other envelope components to strengthen the stratum corneum and maintain tissue resilience. Its expression is coordinated within epidermal differentiation programs involving keratinization and cornification pathways, including transcriptional networks that regulate late differentiation genes. Dysregulated LOR expression or altered cornified envelope assembly is associated with impaired barrier function and has been linked to inherited and inflammatory skin disorders, making it relevant for studies of epidermal homeostasis and disease mechanisms.

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

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

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