ROR2 CRISPR Activation Plasmid (h): sc-401324-ACT

ROR2 encodes receptor tyrosine kinase-like orphan receptor 2, a single-pass transmembrane protein that functions primarily as a noncanonical WNT co-receptor, with strong coupling to WNT5A signaling. ROR2 regulates cytoskeletal remodeling, planar cell polarity, cell migration, and developmental patterning through pathways that intersect with Rho GTPases, JNK signaling, and β-catenin–independent transcriptional programs. In human biology, altered ROR2 expression or signaling is associated with aberrant morphogenesis and dysregulated cell motility, making it relevant to studies of skeletal development, tissue organization, and invasive cellular phenotypes. ROR2 is therefore widely used as a mechanistic entry point for dissecting noncanonical WNT pathway wiring and context-dependent receptor signaling.

ROR2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ROR2 expression without altering the underlying DNA sequence.

ROR2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ROR2 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 ROR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ROR2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ROR2 locus and enabling the study of ROR2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ROR2 pathway restoration in tumor cells with silenced or reduced ROR2 expression.

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