hnRNP A1 CRISPR Activation Plasmid (m): sc-420895-ACT

Mouse Hnrnpa1 encodes hnRNP A1, an abundant RNA-binding protein that associates with pre-mRNAs to coordinate alternative splicing, mRNA export, stability, and translation. hnRNP A1 also shuttles between nucleus and cytoplasm and contributes to stress granule dynamics and RNA quality control, linking RNA processing to cellular stress responses. Through broad transcriptome regulation, hnRNP A1 influences pathways such as spliceosome function, DNA damage responses, and growth-related signaling programs. Dysregulation of hnRNP A1 expression or localization has been associated with altered splicing patterns and perturbed RNA metabolism relevant to neurodegeneration and cancer biology, making it a widely used node for mechanistic studies of post-transcriptional regulation.

hnRNP A1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Hnrnpa1 expression without altering the underlying DNA sequence.

hnRNP A1 CRISPR Activation Plasmid (m) 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.