p130 CRISPR Activation Plasmid (h): sc-400721-ACT

Human RBL2 encodes the retinoblastoma-like protein p130 (RB2), a pocket protein that partners with E2F transcription factors to enforce G0/G1 arrest and restrain S-phase gene expression. p130 is a key effector of the DREAM complex, coordinating cell-cycle quiescence programs and linking cyclin-dependent kinase signaling to transcriptional repression. Through regulation of proliferation, differentiation, and senescence-associated gene networks, altered RBL2/p130 activity is frequently examined in contexts of cell-cycle dysregulation and genome stability. Dysbalanced p130–E2F control is relevant to mechanistic studies of oncogenic signaling and tumor suppressor pathway perturbations in human cells.

p130 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RBL2 expression without altering the underlying DNA sequence.

p130 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RBL2 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 RBL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p130 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RBL2 locus and enabling the study of p130-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p130 pathway restoration in tumor cells with silenced or reduced RBL2 expression.

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