ALPPL2 CRISPR Activation Plasmid (h): sc-401492-ACT

ALPPL2 encodes alkaline phosphatase, placental-like 2, a glycosylphosphatidylinositol (GPI)-anchored ectoenzyme expressed at the cell surface where it hydrolyzes phosphate monoesters and can modulate extracellular nucleotide and phosphate availability. Through its membrane localization and enzymatic activity, ALPPL2 is linked to processes including epithelial differentiation, membrane microdomain organization, and regulation of signaling environments at the plasma membrane. Altered expression of placental-like alkaline phosphatases has been reported in contexts of cellular transformation and lineage state changes, supporting the use of ALPPL2 as a functional marker in studies of tumor biology and cell identity. Its restricted physiological expression pattern relative to ubiquitous alkaline phosphatases makes ALPPL2 relevant for investigating cancer-associated antigen expression programs and phosphatase-dependent remodeling of the extracellular milieu.

ALPPL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ALPPL2 expression without altering the underlying DNA sequence.

ALPPL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ALPPL2 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 ALPPL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ALPPL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ALPPL2 locus and enabling the study of ALPPL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ALPPL2 pathway restoration in tumor cells with silenced or reduced ALPPL2 expression.

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