hnRNP A/B CRISPR Activation Plasmid (h): sc-403454-ACT

HNRNPAB encodes the human heterogeneous nuclear ribonucleoprotein A/B (hnRNP A/B), an RNA-binding protein that associates with nascent transcripts to regulate pre-mRNA splicing, alternative polyadenylation, mRNA stability, and nucleocytoplasmic transport. hnRNP A/B participates in ribonucleoprotein complex assembly and coordinates post-transcriptional gene regulation programs that shape cell-cycle progression and stress-responsive transcriptional outputs. Dysregulation of hnRNP family members, including altered HNRNPAB expression or splicing patterns, has been linked to aberrant RNA processing networks observed in multiple disease contexts, particularly proliferative and neurodegeneration-associated phenotypes. As a result, HNRNPAB is frequently studied in pathways governing RNA metabolism, genome-wide splicing control, and transcriptome remodeling.

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

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

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