OLFML3 CRISPR Activation Plasmid (h): sc-404425-ACT

OLFML3 (olfactomedin-like 3) encodes a secreted extracellular matrix–associated glycoprotein implicated in remodeling of the tissue microenvironment and regulation of cell–matrix interactions. In human cells, OLFML3 has been linked to pathways governing angiogenic signaling, cellular adhesion, and stromal–immune crosstalk, with reported roles in vascular development and extracellular matrix organization. Altered OLFML3 expression has been observed across multiple disease contexts, including cancer-associated stroma and inflammatory settings, supporting its utility as a mechanistic node for studying microenvironment-driven phenotypes. Experimental modulation of OLFML3 is therefore relevant for interrogating how secreted ECM regulators influence migration, endothelial behavior, and signaling networks in complex cultures and co-culture systems.

OLFML3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OLFML3 expression without altering the underlying DNA sequence.

OLFML3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OLFML3 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 OLFML3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OLFML3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OLFML3 locus and enabling the study of OLFML3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OLFML3 pathway restoration in tumor cells with silenced or reduced OLFML3 expression.

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