Hck CRISPR Activation Plasmid (h): sc-401507-ACT

Human HCK encodes Hck, a Src family non-receptor tyrosine kinase expressed predominantly in myeloid lineages where it couples immunoreceptor and integrin signals to downstream phosphorylation cascades. Hck regulates cytoskeletal remodeling, adhesion, chemotaxis, phagocytosis, and inflammatory mediator production through pathways that intersect with PI3K–AKT, MAPK/ERK, and STAT signaling. In hematopoietic and immune contexts, dysregulated Hck activity has been linked to altered innate immune responses and aberrant signaling networks observed in leukemias and other inflammation-associated pathologies. As a proximal kinase in receptor-driven signaling, HCK is frequently studied to dissect stimulus-dependent phosphorylation programs and feedback control in myeloid cells.

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

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

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