ROR1 CRISPR Activation Plasmid (h): sc-401841-ACT

Human ROR1 (receptor tyrosine kinase-like orphan receptor 1) is a single-pass transmembrane receptor of the ROR family implicated in non-canonical Wnt signaling, particularly WNT5A-driven pathways that regulate cytoskeletal dynamics, polarity, and directed migration. ROR1 functions as a context-dependent signaling scaffold that modulates downstream nodes such as Rho GTPases and MAPK-associated programs, influencing cell survival and motility. In adult tissues ROR1 is generally low, while elevated expression is reported in multiple malignancies and is associated with invasive phenotypes and altered differentiation states. These properties make ROR1 a useful target for studying developmental signaling reactivation, epithelial–mesenchymal plasticity, and tumor cell signaling dependencies.

ROR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ROR1 expression without altering the underlying DNA sequence.

ROR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ROR1 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 ROR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ROR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ROR1 locus and enabling the study of ROR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ROR1 pathway restoration in tumor cells with silenced or reduced ROR1 expression.

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