PTEN CRISPR Activation Plasmid (m): sc-422475-ACT

Mouse Pten encodes PTEN, a lipid and protein phosphatase that counteracts PI3K signaling by dephosphorylating PIP3, thereby restraining AKT/mTOR pathway activation and shaping cellular growth, metabolism, and survival programs. PTEN also contributes to genome stability, cell-cycle control, and cell polarity through cytoplasmic and nuclear functions that influence DNA damage responses and chromatin regulation. Disruption or reduced expression of PTEN is broadly linked to dysregulated proliferative signaling and altered differentiation states across tissues, making it a central node in studies of oncogenic pathway rewiring. In murine models, Pten dosage effects are frequently used to interrogate pathway crosstalk with receptor tyrosine kinases, MAPK signaling, and immune microenvironmental cues.

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

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

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