CRTAP CRISPR Activation Plasmid (h): sc-405828-ACT

CRTAP (cartilage associated protein) is an endoplasmic reticulum–resident component of the prolyl 3-hydroxylation complex that collaborates with P3H1/LEPRE1 and PPIB/cyclophilin B to modify and chaperone nascent fibrillar collagens. Through regulation of collagen post-translational processing, CRTAP supports proper triple-helix folding, secretion, and extracellular matrix assembly, linking it to collagen biosynthesis and ER proteostasis pathways. Disruption or reduced function of CRTAP perturbs connective tissue homeostasis and is implicated in heritable skeletal dysplasias, including recessive forms of osteogenesis imperfecta. As a result, CRTAP is widely studied in matrix biology, osteoblast and chondrocyte differentiation, and mechanisms of ER stress associated with aberrant collagen maturation.

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

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

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