PiT1 CRISPR Activation Plasmid (m): sc-422988-ACT

Slc20a1 encodes PiT1 (SLC20A1), a sodium-dependent inorganic phosphate transporter that supports phosphate uptake required for nucleotide synthesis, phospholipid metabolism, and energy homeostasis. Beyond transport, PiT1 contributes to signaling networks that couple phosphate availability to cell proliferation, survival, and differentiation, with links to MAPK and other growth-associated pathways. In mouse systems, Slc20a1 activity is frequently examined in contexts where phosphate handling influences osteogenic and vascular calcification programs, as well as developmental and tissue remodeling processes. Dysregulated phosphate transport and downstream signaling are therefore relevant to mechanistic studies of mineralization, metabolic stress responses, and proliferative phenotypes.

PiT1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc20a1 expression without altering the underlying DNA sequence.

PiT1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slc20a1 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 Slc20a1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PiT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slc20a1 locus and enabling the study of PiT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PiT1 pathway restoration in tumor cells with silenced or reduced Slc20a1 expression.

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