hnRNP E1 CRISPR Activation Plasmid (h): sc-401557-ACT

PCBP1 encodes heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1), an RNA- and DNA-binding factor that coordinates post-transcriptional gene regulation by controlling mRNA stability, alternative splicing, transport, and translation. hnRNP E1 recognizes C-rich sequence elements within target transcripts and participates in ribonucleoprotein complexes that couple signaling inputs to regulated protein synthesis. Through these activities, PCBP1 contributes to cellular processes including differentiation, stress responses, and maintenance of proteome homeostasis. Altered PCBP1/hnRNP E1 expression or function has been associated with dysregulated gene expression programs observed in cancer biology and other disorders linked to aberrant RNA processing.

hnRNP E1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCBP1 expression without altering the underlying DNA sequence.

hnRNP E1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCBP1 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 PCBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous hnRNP E1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCBP1 locus and enabling the study of hnRNP E1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of hnRNP E1 pathway restoration in tumor cells with silenced or reduced PCBP1 expression.

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