Pellino 2 CRISPR Activation Plasmid (h): sc-402305-ACT

Human PELI2 encodes Pellino 2, an E3 ubiquitin ligase that helps couple pattern-recognition receptor signaling to downstream inflammatory and antiviral responses. Pellino 2 participates in ubiquitin-dependent regulation of adaptor and kinase complexes within Toll-like receptor and interleukin-1 receptor pathways, shaping activation of NF-κB and MAPK signaling and influencing cytokine production. By modulating protein turnover and signal amplitude, PELI2 contributes to innate immune homeostasis and cellular stress responses relevant to inflammatory signaling dysregulation. Altered regulation of ubiquitination and innate immune pathways involving Pellino 2 has been studied in the context of immune-mediated disease mechanisms and host–pathogen interactions.

Pellino 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PELI2 expression without altering the underlying DNA sequence.

Pellino 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PELI2 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 PELI2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pellino 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PELI2 locus and enabling the study of Pellino 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pellino 2 pathway restoration in tumor cells with silenced or reduced PELI2 expression.

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