ARID3A CRISPR Activation Plasmid (h2): sc-404552-ACT-2

Human ARID3A encodes an AT-rich interaction domain–containing DNA-binding transcription factor that regulates chromatin accessibility and gene expression programs involved in lineage specification, particularly during B cell development and immunoglobulin gene regulation. ARID3A participates in transcriptional control networks governing cell cycle progression, differentiation, and epigenetic remodeling through interactions with promoter/enhancer elements and coregulatory complexes. Dysregulated ARID3A expression or activity has been linked to aberrant hematopoietic differentiation and immune signaling, and is frequently studied in the context of oncogenic transcriptional programs and autoimmunity-associated gene signatures. Gene editing of ARID3A in human cells enables mechanistic dissection of transcription factor binding dependencies, enhancer function, and downstream pathway rewiring using assays such as RNA-seq, ATAC-seq, ChIP-seq, and functional differentiation models.

ARID3A CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous ARID3A expression without altering the underlying DNA sequence.

ARID3A CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARID3A 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 ARID3A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARID3A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARID3A locus and enabling the study of ARID3A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARID3A pathway restoration in tumor cells with silenced or reduced ARID3A expression.

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