ASPP1 CRISPR Activation Plasmid (m): sc-423473-ACT

Mouse Ppp1r13b encodes ASPP1, a conserved regulator of the p53 family that enhances sequence-specific transcriptional programs controlling apoptosis, cell-cycle arrest, and DNA damage responses. ASPP1 modulates the balance between pro-survival and pro-death signaling by influencing p53, p63, and p73 target gene selection and cooperates with stress-responsive pathways that shape epithelial homeostasis and differentiation. Through these functions, altered ASPP1 activity is relevant to experimental models of tumor suppression, genomic instability, and context-dependent remodeling of cell fate decisions. Ppp1r13b is therefore a useful node for dissecting transcriptional control mechanisms that couple cellular stress sensing to programmed cell death.

ASPP1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ppp1r13b expression without altering the underlying DNA sequence.

ASPP1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ppp1r13b 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 Ppp1r13b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ASPP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ppp1r13b locus and enabling the study of ASPP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ASPP1 pathway restoration in tumor cells with silenced or reduced Ppp1r13b expression.

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