RHAMM CRISPR Activation Plasmid (h): sc-402431-ACT

HMMR encodes RHAMM (hyaluronan-mediated motility receptor), a multifunctional protein that binds hyaluronan and interfaces with cytoskeletal regulators to coordinate cell motility, polarity, and extracellular matrix–driven signaling. RHAMM localizes to the cell surface, cytoplasm, and mitotic spindle, where it contributes to microtubule dynamics, centrosome function, and mitotic progression through pathways including ERK/MAPK signaling and spindle assembly control. Dysregulated HMMR expression has been associated with altered migration and proliferation phenotypes and is frequently studied in the context of tumor cell invasion, metastasis-associated programs, and aberrant cell cycle regulation. As a result, RHAMM serves as a useful node for dissecting hyaluronan–ECM signaling, cytoskeletal remodeling, and mitosis-linked stress responses in human cell models.

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

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

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