PHLPPL CRISPR Activation Plasmid (m): sc-434083-ACT

Mouse Phlpp2 encodes PHLPPL, a PP2C-family Ser/Thr phosphatase that attenuates growth factor signaling by dephosphorylating AGC kinases, most notably AKT at Ser473, thereby constraining PI3K–AKT pathway amplitude and downstream mTOR-linked metabolic and survival programs. Through negative regulation of kinase phosphorylation, PHLPPL helps shape cellular decisions involving proliferation, apoptosis, and stress responses, and can influence feedback control across receptor tyrosine kinase signaling networks. Altered PHLPP/PHLPPL activity has been associated in the literature with dysregulated signaling homeostasis relevant to oncogenic transformation, insulin/energy metabolism, and inflammation-related phenotypes, making Phlpp2 a useful node for mechanistic studies of pathway buffering. As a phosphatase that counterbalances kinase-driven cascades, PHLPPL is also relevant for dissecting signal-duration effects and adaptive resistance mechanisms in cell models.

PHLPPL CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Phlpp2 expression without altering the underlying DNA sequence.

PHLPPL CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Phlpp2 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 Phlpp2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PHLPPL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Phlpp2 locus and enabling the study of PHLPPL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PHLPPL pathway restoration in tumor cells with silenced or reduced Phlpp2 expression.

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