Gab 1 CRISPR Activation Plasmid (h): sc-401108-ACT

Human GAB1 encodes Gab 1, a scaffolding adaptor that integrates signals from receptor tyrosine kinases and cytokine receptors to coordinate downstream cascades controlling proliferation, survival, migration, and differentiation. Upon tyrosine phosphorylation, Gab 1 recruits SH2-domain effectors such as PTPN11/SHP2 and the p85 subunit of PI3K, linking upstream receptors to MAPK/ERK and PI3K–AKT signaling and associated cytoskeletal and vesicular trafficking processes. Gab 1-dependent signaling contributes to regulation of epithelial morphogenesis, immune cell activation, and growth factor responses. Dysregulated GAB1 expression or phosphorylation has been associated with aberrant RTK pathway output observed across multiple disease contexts, supporting its use as a mechanistic node in pathway and network studies.

Gab 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GAB1 expression without altering the underlying DNA sequence.

Gab 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GAB1 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 GAB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Gab 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GAB1 locus and enabling the study of Gab 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Gab 1 pathway restoration in tumor cells with silenced or reduced GAB1 expression.

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