MOB1A CRISPR Activation Plasmid (h): sc-407966-ACT

Human MOB1A encodes MOB kinase activator 1A, a core cofactor of the Hippo signaling cascade that binds and activates LATS1/2 kinases to restrain YAP/TAZ-dependent transcriptional programs. Through this pathway, MOB1A contributes to control of cell cycle progression, apoptosis, epithelial polarity, and contact inhibition, integrating cues from cytoskeletal dynamics and cell–cell junctions. Altered Hippo pathway regulation and dysregulated YAP/TAZ activity are broadly implicated in oncogenic transformation, metastasis, and aberrant tissue growth, making MOB1A a relevant node for mechanistic studies of proliferation and differentiation. MOB1A function is also studied in contexts of stress responses and organ size homeostasis where precise tuning of transcriptional outputs is required.

MOB1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MOB1A expression without altering the underlying DNA sequence.

MOB1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MOB1A 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 MOB1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MOB1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MOB1A locus and enabling the study of MOB1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MOB1A pathway restoration in tumor cells with silenced or reduced MOB1A expression.

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