Histone cluster 1 H1E CRISPR Activation Plasmid (h): sc-402775-ACT

HIST1H1E encodes histone cluster 1 H1E, a linker histone H1 family member that binds nucleosomal DNA at the entry/exit sites to promote higher-order chromatin compaction and regulate genome accessibility. By influencing nucleosome spacing and chromatin fiber organization, H1E contributes to transcriptional control, DNA replication timing, and DNA damage response signaling. Altered H1 stoichiometry and H1E-associated chromatin states are studied in the context of epigenetic reprogramming, differentiation, and genome stability, processes frequently perturbed in cancer and neurodevelopmental disorders. As a chromatin architectural protein, H1E is also relevant to investigations of replication stress, heterochromatin formation, and transcription factor occupancy across the genome.

Histone cluster 1 H1E CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HIST1H1E expression without altering the underlying DNA sequence.

Histone cluster 1 H1E CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HIST1H1E 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 HIST1H1E transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Histone cluster 1 H1E expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HIST1H1E locus and enabling the study of Histone cluster 1 H1E-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Histone cluster 1 H1E pathway restoration in tumor cells with silenced or reduced HIST1H1E expression.

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