kpm CRISPR Activation Plasmid (h): sc-404134-ACT

Human LATS2 encodes a core serine/threonine kinase of the Hippo signaling pathway that constrains cell proliferation and promotes apoptosis by regulating YAP/TAZ transcriptional co-activators. Through phosphorylation-dependent control of nuclear YAP/TAZ activity, LATS2 helps maintain tissue homeostasis, contact inhibition, and cell-cycle checkpoint integrity, with additional links to centrosome biology and stress responses. Altered LATS2 function or Hippo pathway dysregulation is frequently associated with abnormal growth control and genome instability in diverse disease-relevant contexts, making it a widely studied node in tumor suppressor networks. As a kinase positioned at pathway convergence, LATS2 is also used to interrogate crosstalk between Hippo signaling, cytoskeletal dynamics, and transcriptional programs governing epithelial-to-mesenchymal features.

kpm CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LATS2 expression without altering the underlying DNA sequence.

kpm CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LATS2 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 LATS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous kpm expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LATS2 locus and enabling the study of kpm-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of kpm pathway restoration in tumor cells with silenced or reduced LATS2 expression.

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