B-ATF CRISPR Activation Plasmid (h): sc-401553-ACT

BATF (B-ATF) is a basic leucine zipper transcription factor that heterodimerizes with JUN family proteins to bind AP-1–like regulatory elements and shape lineage-specific gene expression programs. In human immune cells, BATF is a key regulator of T cell differentiation and effector function, including pathways controlling cytokine production, T follicular helper cell biology, and adaptive immune activation. By cooperating with IRF family members at composite DNA motifs, BATF influences chromatin accessibility and transcriptional networks that coordinate immune signaling and cellular stress responses. Dysregulated BATF activity has been associated with altered immune phenotypes and has been implicated in inflammatory disease mechanisms and oncogenic transcriptional states in lymphoid contexts, making it a useful node for mechanistic studies of transcriptional control.

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

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

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