ASF1A CRISPR Activation Plasmid (h): sc-404071-ACT

ASF1A encodes anti-silencing function 1A, a conserved histone H3–H4 chaperone that coordinates nucleosome assembly and disassembly during DNA replication, transcription, and DNA damage repair. By partnering with chromatin regulators such as CAF-1 and HIRA, ASF1A supports replication-coupled chromatin reassembly, histone supply dynamics, and maintenance of epigenetic states across the cell cycle. Altered ASF1A activity can perturb genome stability programs and chromatin accessibility, linking this factor to dysregulated proliferation and stress responses observed in cancer and other diseases with chromatin remodeling defects. As a central node in chromatin homeostasis, ASF1A is frequently studied in pathways governing replication stress, checkpoint signaling, and transcriptional reprogramming.

ASF1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ASF1A expression without altering the underlying DNA sequence.

ASF1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ASF1A 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 ASF1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ASF1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ASF1A locus and enabling the study of ASF1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ASF1A pathway restoration in tumor cells with silenced or reduced ASF1A expression.

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