CD5L CRISPR Activation Plasmid (h): sc-404865-ACT

CD5L (CD5 molecule-like) is a secreted scavenger receptor cysteine-rich protein predominantly produced by macrophages that modulates innate immune homeostasis and inflammation. It binds diverse lipid and microbial ligands and influences processes including macrophage polarization, apoptotic cell handling, and lipid metabolism, integrating with pathways linked to cytokine signaling and tissue remodeling. CD5L has been associated with immune-mediated pathology and metabolic inflammation, and altered expression has been reported across contexts such as atherosclerotic lesions, chronic inflammatory states, and tumor-associated macrophage biology. These features make CD5L a useful target for dissecting macrophage-driven microenvironmental cues and lipid-immune crosstalk in human cellular models.

CD5L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD5L expression without altering the underlying DNA sequence.

CD5L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD5L 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 CD5L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD5L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD5L locus and enabling the study of CD5L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD5L pathway restoration in tumor cells with silenced or reduced CD5L expression.

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