COL11A1 CRISPR Activation Plasmid (h): sc-403512-ACT

COL11A1 encodes the α1 chain of type XI collagen, a quantitatively minor but structurally critical fibrillar collagen that regulates collagen fibril nucleation, diameter, and extracellular matrix (ECM) organization in connective tissues. COL11A1 integrates into heterotypic fibrils with collagens II and IX, supporting cartilage biomechanics and influencing cell–matrix adhesion, migration, and mechanotransduction pathways that shape tissue architecture. Altered COL11A1 expression and ECM remodeling are associated with developmental skeletal phenotypes and have been linked to tumor-associated stromal programs, where matrix composition can modulate invasion-related signaling. As a matrix component, COL11A1 is frequently studied in the context of ECM assembly, chondrogenesis, and microenvironment-dependent regulation of cell behavior.

COL11A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL11A1 expression without altering the underlying DNA sequence.

COL11A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL11A1 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 COL11A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL11A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL11A1 locus and enabling the study of COL11A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL11A1 pathway restoration in tumor cells with silenced or reduced COL11A1 expression.

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