CKR-9 CRISPR Activation Plasmid (m): sc-419702-ACT

Mouse Ccr9 encodes the chemokine receptor CKR-9 (CCR9), a GPCR that preferentially binds CCL25 to guide leukocyte trafficking, especially thymocyte development and gut-homing of T cell subsets. CCR9 signaling engages canonical chemokine pathways involving Gαi-dependent responses, intracellular calcium flux, PI3K–AKT, and MAPK cascades that coordinate chemotaxis, adhesion, and tissue localization. Altered CCR9–CCL25 axis activity has been linked to dysregulated mucosal immunity and inflammatory processes, and it is frequently studied in contexts such as intestinal immune homeostasis and lymphocyte migration. In mouse models, Ccr9 expression is also used as a marker and functional determinant of tissue-resident and migratory immune cell programs relevant to immune-mediated disease mechanisms.

CKR-9 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ccr9 expression without altering the underlying DNA sequence.

CKR-9 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ccr9 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 Ccr9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CKR-9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ccr9 locus and enabling the study of CKR-9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CKR-9 pathway restoration in tumor cells with silenced or reduced Ccr9 expression.

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