HOOK1 CRISPR Activation Plasmid (h): sc-404872-ACT

Human HOOK1 encodes a microtubule-binding adaptor protein that links dynein–dynactin motors to endosomal and organelle cargo, supporting minus-end-directed transport and spatial organization of membrane trafficking. Through its role in microtubule-dependent dynamics, HOOK1 contributes to centrosome-associated processes, ciliary assembly and maintenance, and coordination of endosome-to-Golgi trafficking pathways. Perturbation of HOOK1-regulated transport can influence cell polarity, mitotic progression, and signal compartmentalization, making it relevant to studies of neurodevelopmental biology, ciliopathy-related mechanisms, and dysregulated intracellular trafficking observed in diverse disease contexts. Altered HOOK1 expression or function has also been investigated in relation to tumor cell behavior and genome stability pathways that depend on proper cytoskeletal organization.

HOOK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOOK1 expression without altering the underlying DNA sequence.

HOOK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOOK1 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 HOOK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HOOK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOOK1 locus and enabling the study of HOOK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HOOK1 pathway restoration in tumor cells with silenced or reduced HOOK1 expression.

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