hnRNP K CRISPR Activation Plasmid (h): sc-417266-ACT

HNRNPK encodes heterogeneous nuclear ribonucleoprotein K (hnRNP K), a multifunctional nucleic acid–binding protein that coordinates transcription, pre-mRNA splicing, mRNA stability, and translation through interactions with poly(C) RNA/DNA elements and diverse signaling partners. hnRNP K integrates cues from pathways such as MAPK/ERK and DNA damage response networks to modulate chromatin-associated gene expression programs and stress-adaptive RNA processing. By acting as a scaffold for ribonucleoprotein complexes, it influences cell-cycle progression, apoptosis, and differentiation. Dysregulated HNRNPK expression or function has been associated with altered oncogenic and neurodevelopmental gene regulation, making it a useful node for mechanistic studies of RNA biology and signal-dependent transcriptional control.

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

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

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