MyD88 CRISPR Activation Plasmid (h): sc-417166-ACT

MYD88 encodes MyD88, a central adaptor protein for Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) signaling that couples receptor engagement to downstream NF-κB and MAPK activation. Through its TIR and death domains, MyD88 coordinates assembly of the myddosome complex with IRAK kinases and TRAF6, promoting inflammatory gene expression and innate immune cell activation. Dysregulated MYD88 signaling is implicated in aberrant cytokine production, chronic inflammation, and oncogenic signaling programs in select hematologic malignancies, making it a key node for mechanistic studies of immune signaling. MYD88 also interfaces with cell survival and stress-response pathways, supporting research into context-dependent crosstalk between innate sensing and transcriptional control.

MyD88 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYD88 expression without altering the underlying DNA sequence.

MyD88 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYD88 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 MYD88 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MyD88 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYD88 locus and enabling the study of MyD88-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MyD88 pathway restoration in tumor cells with silenced or reduced MYD88 expression.

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