LMP7 CRISPR Activation Plasmid (m): sc-421450-ACT

Psmb8 encodes the mouse immunoproteasome catalytic subunit LMP7 (β5i), a key component of the IFN-γ–inducible 20S proteasome that reshapes proteolytic specificity during inflammation. LMP7 promotes generation of MHC class I peptides and supports antigen processing and presentation, linking proteostasis to adaptive immune surveillance. By influencing NF-κB–associated inflammatory programs and cellular responses to cytokine stress, Psmb8 activity impacts immune cell differentiation and activation states. Dysregulated immunoproteasome function has been connected to aberrant antigen presentation and chronic inflammatory phenotypes, making Psmb8 a relevant target for immunology and proteostasis research in mouse models.

LMP7 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Psmb8 expression without altering the underlying DNA sequence.

LMP7 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Psmb8 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 Psmb8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LMP7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Psmb8 locus and enabling the study of LMP7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LMP7 pathway restoration in tumor cells with silenced or reduced Psmb8 expression.

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