hnRNP C1/C2 CRISPR Activation Plasmid (h): sc-400993-ACT

HNRNPC encodes heterogeneous nuclear ribonucleoprotein C1/C2, abundant nuclear RNA-binding proteins that assemble on nascent pre-mRNA to regulate splice site selection, alternative splicing fidelity, and mRNA maturation. hnRNP C1/C2 participates in co‑transcriptional RNA processing, impacts mRNA export and stability, and helps maintain transcriptome integrity by coordinating ribonucleoprotein complex formation. Through these functions it interfaces with core gene expression programs, including RNA metabolism pathways and cell-cycle–linked transcriptional responses. Dysregulated HNRNPC expression or hnRNP C1/C2 activity has been associated with altered splicing landscapes and aberrant RNA processing observed across multiple disease contexts, supporting its use as a mechanistic node in studies of RNA-driven phenotypes.

hnRNP C1/C2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HNRNPC expression without altering the underlying DNA sequence.

hnRNP C1/C2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HNRNPC 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 HNRNPC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous hnRNP C1/C2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HNRNPC locus and enabling the study of hnRNP C1/C2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of hnRNP C1/C2 pathway restoration in tumor cells with silenced or reduced HNRNPC expression.

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