ATP11B CRISPR Activation Plasmid (h): sc-411692-ACT

ATP11B encodes a P4-ATPase phospholipid flippase that helps maintain plasma membrane and endomembrane lipid asymmetry by translocating aminophospholipids across bilayers in an ATP-dependent manner. Through regulation of membrane curvature and leaflet composition, ATP11B contributes to vesicle budding, endocytic recycling, and Golgi-associated trafficking processes that shape protein sorting and cellular polarity. Altered phospholipid transport and membrane dynamics involving ATP11B have been linked in the literature to changes in drug transport phenotypes and stress responses in tumor cell models, supporting investigation in mechanisms of chemoresistance and intracellular trafficking. Its activity also intersects with pathways controlling membrane homeostasis, apoptosis signaling linked to lipid scrambling, and organelle function.

ATP11B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATP11B expression without altering the underlying DNA sequence.

ATP11B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATP11B 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 ATP11B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATP11B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATP11B locus and enabling the study of ATP11B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATP11B pathway restoration in tumor cells with silenced or reduced ATP11B expression.

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