PTBP-2 CRISPR Activation Plasmid (h): sc-404312-ACT

Human PTBP2 encodes the RNA-binding protein PTBP-2, a neuron-enriched regulator of alternative pre-mRNA splicing that influences exon inclusion, mRNA stability, and translation during neurogenesis and synaptic maturation. PTBP-2 coordinates splice-site selection across transcripts involved in neuronal differentiation, cytoskeletal remodeling, and activity-dependent gene expression, intersecting with post-transcriptional regulatory networks that shape cell identity. Altered PTBP2 expression or splicing programs have been associated with dysregulated neuronal development and aberrant RNA processing patterns observed in neurodevelopmental and neurodegenerative disease contexts. As a modulator of transcript isoform balance, PTBP-2 is widely studied to link splicing decisions to neural lineage commitment and neuronal function.

PTBP-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTBP2 expression without altering the underlying DNA sequence.

PTBP-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTBP2 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 PTBP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PTBP-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTBP2 locus and enabling the study of PTBP-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PTBP-2 pathway restoration in tumor cells with silenced or reduced PTBP2 expression.

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