DOCK 11 CRISPR Activation Plasmid (h): sc-406204-ACT

DOCK11 (dedicator of cytokinesis 11) encodes a Rho family guanine nucleotide exchange factor that preferentially activates Cdc42 to coordinate actin cytoskeleton remodeling, cell polarity, and directional migration. Through regulation of small GTPase signaling, DOCK11 contributes to immune cell trafficking, adhesion dynamics, and processes linked to endocytic and membrane remodeling events. Altered DOCK11 activity has been associated with immune dysregulation phenotypes and has been studied in the context of hematologic and lymphoid biology, where Rho GTPase-dependent signaling influences proliferation and cellular interactions. As a human gene, DOCK11 is of interest for dissecting Cdc42-driven pathways that couple receptor cues to cytoskeletal organization and transcriptional programs.

DOCK 11 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DOCK11 expression without altering the underlying DNA sequence.

DOCK 11 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DOCK11 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 DOCK11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DOCK 11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DOCK11 locus and enabling the study of DOCK 11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DOCK 11 pathway restoration in tumor cells with silenced or reduced DOCK11 expression.

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