hnRNP M CRISPR Activation Plasmid (h): sc-401670-ACT

HNRNPM encodes heterogeneous nuclear ribonucleoprotein M (hnRNP M), an RNA-binding protein that associates with nascent transcripts to regulate pre-mRNA processing, alternative splicing, and mRNA maturation. hnRNP M participates in spliceosome-related RNA metabolism and influences transcript isoform selection that can affect cell-state programs such as proliferation, differentiation, and stress-adaptive gene expression. Dysregulation of HNRNPM-dependent splicing networks has been linked to altered epithelial–mesenchymal transition and aberrant RNA processing signatures observed across multiple disease contexts, making it a useful node for studying pathway-level consequences of splicing factor perturbation. Experimental modulation of hnRNP M levels can help map isoform-switching events, RNA–protein interaction dependencies, and downstream signaling changes driven by transcriptome remodeling.

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

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

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