DOCK 180 CRISPR Activation Plasmid (h): sc-401387-ACT

Human DOCK1 (DOCK 180) is an atypical guanine nucleotide exchange factor that activates Rac-family GTPases to coordinate actin cytoskeleton remodeling, cell adhesion, and directed migration. Through signaling nodes downstream of integrins, receptor tyrosine kinases, and chemokine receptors, DOCK 180 supports processes such as lamellipodia formation, macropinocytosis, and engulfment of apoptotic cells. DOCK1-dependent Rac signaling interfaces with pathways that regulate epithelial–mesenchymal transitions, invasion-associated motility programs, and innate immune cell trafficking. Dysregulated DOCK1 activity or expression has been associated with inflammatory phenotypes and tumor cell dissemination-relevant behaviors, making it a useful target for mechanistic studies in cancer biology and immunology.

DOCK 180 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DOCK1 expression without altering the underlying DNA sequence.

DOCK 180 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DOCK1 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 DOCK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DOCK 180 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DOCK1 locus and enabling the study of DOCK 180-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DOCK 180 pathway restoration in tumor cells with silenced or reduced DOCK1 expression.

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