HBXIP CRISPR Activation Plasmid (h): sc-405228-ACT

LAMTOR5 encodes HBXIP (hepatitis B virus X-interacting protein), an endosomal/lysosomal adaptor best known as a component of the Ragulator complex that helps couple amino acid sensing to mTORC1 activation at the lysosomal surface. Through regulation of nutrient-responsive signaling, HBXIP influences cellular growth, autophagy, and metabolic adaptation, and has also been linked to control of cell-cycle progression and transcriptional programs. Dysregulated HBXIP expression or pathway engagement has been associated with altered proliferative signaling and stress responses reported across multiple cancer-relevant contexts. As a result, LAMTOR5/HBXIP is commonly studied in mechanisms connecting lysosome-centric signaling hubs to oncogenic metabolism, proteostasis, and intracellular trafficking.

HBXIP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAMTOR5 expression without altering the underlying DNA sequence.

HBXIP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAMTOR5 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 LAMTOR5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HBXIP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAMTOR5 locus and enabling the study of HBXIP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HBXIP pathway restoration in tumor cells with silenced or reduced LAMTOR5 expression.

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