Rac 2 CRISPR Activation Plasmid (h): sc-401157-ACT

RAC2 encodes Rac2, a hematopoietic-enriched Rho family small GTPase that cycles between GDP- and GTP-bound states to coordinate actin cytoskeletal remodeling, cell polarity, and membrane trafficking. Rac2 functions downstream of immunoreceptor, integrin, and chemokine signaling to regulate NADPH oxidase activation, reactive oxygen species production, and phagocyte antimicrobial responses, while also influencing MAPK- and PI3K-dependent signaling outputs. In leukocytes, Rac2 supports chemotaxis, adhesion, degranulation, and immune synapse dynamics, linking cytoskeletal control to inflammatory and innate immune pathways. Dysregulated RAC2 activity or expression has been associated with altered host defense and immune homeostasis, making it relevant for studying mechanisms of immunodeficiency, inflammatory phenotypes, and hematologic cell signaling.

Rac 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAC2 expression without altering the underlying DNA sequence.

Rac 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAC2 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 RAC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rac 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAC2 locus and enabling the study of Rac 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rac 2 pathway restoration in tumor cells with silenced or reduced RAC2 expression.

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