PKC lambda/iota CRISPR Activation Plasmid (h): sc-402111-ACT

PRKCI encodes the atypical protein kinase C isoform PKC lambda/iota, a serine/threonine kinase that functions within polarity and signaling complexes to regulate epithelial organization, asymmetric cell division, and cytoskeletal dynamics. Acting downstream of phosphoinositide and small GTPase inputs, PRKCI participates in Par3–Par6 polarity signaling and interfaces with pathways controlling cell survival, migration, and junctional integrity. Dysregulated PRKCI activity or expression has been associated with altered polarity programs and proliferative signaling in multiple disease contexts, including cancer-related phenotypes such as invasion and resistance to apoptotic cues. These features make PRKCI a useful target for mechanistic studies of polarity-dependent signaling, stress responses, and cell-state transitions.

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

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

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