hnRNP E2 CRISPR Activation Plasmid (h): sc-402274-ACT

PCBP2 encodes heterogeneous nuclear ribonucleoprotein E2 (hnRNP E2), an RNA-binding protein that recognizes C-rich elements to coordinate post-transcriptional gene regulation. hnRNP E2 influences mRNA splicing, stability, polyadenylation, and translation, thereby shaping protein output in response to cellular cues and stress. It contributes to ribonucleoprotein complex assembly and interacts with networks controlling RNA metabolism and translation initiation. Dysregulated PCBP2/hnRNP E2 activity has been associated with altered gene expression programs in cancer biology, viral RNA utilization, and inflammatory signaling contexts, supporting its study in mechanisms of gene regulation and cell-state control.

hnRNP E2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCBP2 expression without altering the underlying DNA sequence.

hnRNP E2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCBP2 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 PCBP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous hnRNP E2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCBP2 locus and enabling the study of hnRNP E2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of hnRNP E2 pathway restoration in tumor cells with silenced or reduced PCBP2 expression.

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