IRAK-2 CRISPR Activation Plasmid (h): sc-404304-ACT

Human IRAK2 encodes interleukin-1 receptor–associated kinase 2 (IRAK-2), a signaling kinase that couples Toll-like receptor (TLR) and IL-1 receptor engagement to downstream inflammatory transcriptional programs. IRAK-2 functions within the MyD88-dependent pathway to promote assembly and signaling of the IRAK complex, facilitating activation of NF-κB and MAPK cascades and shaping cytokine and chemokine expression. Through these processes, IRAK-2 helps coordinate innate immune responses, with dysregulated signaling linked to chronic inflammation and altered immune homeostasis. Its pathway connectivity makes IRAK2 a useful node for studying stimulus-dependent transcriptional outputs, feedback regulation, and cross-talk between inflammatory and stress-response pathways.

IRAK-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IRAK2 expression without altering the underlying DNA sequence.

IRAK-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IRAK2 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 IRAK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IRAK-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IRAK2 locus and enabling the study of IRAK-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IRAK-2 pathway restoration in tumor cells with silenced or reduced IRAK2 expression.

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