LBP CRISPR Activation Plasmid (h): sc-402651-ACT

Human LBP (lipopolysaccharide binding protein) is a soluble acute-phase pattern recognition molecule that binds bacterial lipopolysaccharide and transfers it to CD14/MD-2/TLR4 complexes, amplifying innate immune detection of Gram-negative bacteria. By facilitating TLR4-dependent NF-κB and MAPK signaling, LBP modulates cytokine production, leukocyte activation, and systemic inflammatory responses. LBP expression is regulated by inflammatory mediators and hepatic acute-phase programs, linking it to host–microbe interactions and endotoxin responsiveness. Dysregulated LBP levels and signaling context have been associated with inflammatory and metabolic phenotypes, making it a useful marker and mechanistic node in studies of barrier dysfunction, infection biology, and immune-driven pathology.

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

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

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