hnRNP A2/B1 CRISPR Activation Plasmid (m): sc-424702-ACT

Mouse Hnrnpa2b1 encodes the RNA-binding protein hnRNP A2/B1, a core component of heterogeneous nuclear ribonucleoprotein complexes that coordinates pre-mRNA splicing, mRNA export, stability, and translation. hnRNP A2/B1 participates in co- and post-transcriptional gene regulation through interactions with spliceosomal factors and coupling of transcription to RNA processing, influencing cell-state programs such as proliferation and differentiation. It has also been implicated in RNA granule biology and stress responses, linking RNA metabolism to proteostasis and subcellular compartmentalization. Dysregulation of hnRNP A2/B1-dependent RNA processing and isoform selection is frequently studied in contexts of neurodegeneration, oncogenic reprogramming, and inflammatory signaling where aberrant RNA regulation contributes to disease-associated phenotypes.

hnRNP A2/B1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Hnrnpa2b1 expression without altering the underlying DNA sequence.

hnRNP A2/B1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Hnrnpa2b1 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 Hnrnpa2b1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous hnRNP A2/B1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Hnrnpa2b1 locus and enabling the study of hnRNP A2/B1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of hnRNP A2/B1 pathway restoration in tumor cells with silenced or reduced Hnrnpa2b1 expression.

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