hnRNP A1 CRISPR Activation Plasmid (h): sc-400704-ACT

HNRNPA1 encodes the human heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), an abundant RNA-binding protein that coordinates pre-mRNA splicing, alternative exon selection, mRNA nuclear export, and translation regulation through interactions with spliceosomal components and RNA transport machinery. hnRNP A1 also participates in telomere maintenance and stress granule dynamics, linking RNA metabolism to cellular stress responses. Dysregulation of HNRNPA1-dependent RNA processing and protein aggregation has been associated with neurodegenerative and neuromuscular phenotypes, and altered hnRNP A1 activity has been reported across multiple cancer contexts where transcript isoform remodeling influences proliferation and survival pathways. As a central regulator of transcriptome plasticity, hnRNP A1 is frequently studied in pathways governing RNA stability, DNA damage response–adjacent gene expression programs, and proteostasis.

hnRNP A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HNRNPA1 expression without altering the underlying DNA sequence.

hnRNP A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HNRNPA1 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 HNRNPA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous hnRNP A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HNRNPA1 locus and enabling the study of hnRNP A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of hnRNP A1 pathway restoration in tumor cells with silenced or reduced HNRNPA1 expression.

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