CAF-1 p150 CRISPR Activation Plasmid (h): sc-402472-ACT

CHAF1A encodes CAF-1 p150, the largest subunit of the chromatin assembly factor-1 complex that deposits histones H3–H4 onto newly synthesized DNA during replication-coupled nucleosome assembly. By coordinating DNA replication with chromatin restoration, CAF-1 p150 supports S-phase progression, replication fork stability, and maintenance of epigenetic states, and it contributes to genome integrity through interactions with PCNA and DNA repair factors. Disruption of CAF-1 function is linked to replication stress, altered heterochromatin organization, and aberrant transcriptional programs, processes frequently studied in the context of proliferative disease biology. CHAF1A activity is therefore relevant to pathways governing DNA damage responses, chromatin dynamics, and cell cycle control.

CAF-1 p150 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHAF1A expression without altering the underlying DNA sequence.

CAF-1 p150 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHAF1A 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 CHAF1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CAF-1 p150 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHAF1A locus and enabling the study of CAF-1 p150-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CAF-1 p150 pathway restoration in tumor cells with silenced or reduced CHAF1A expression.

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