Bcl-11b CRISPR Activation Plasmid (h): sc-403460-ACT

BCL11B encodes the transcription factor Bcl-11b, a C2H2 zinc-finger protein that functions as a lineage-defining regulator of T cell development and thymocyte differentiation. It integrates signals from key developmental pathways, including Notch and TCR-driven transcriptional programs, to control chromatin state and coordinate gene expression networks governing proliferation, survival, and commitment. Dysregulated BCL11B activity has been linked to altered immune cell identity and aberrant transcriptional control in hematologic disease contexts, making it a relevant node for studying gene regulatory circuitry. In human cell models, perturbing Bcl-11b levels is commonly used to interrogate differentiation, enhancer logic, and transcription factor cooperation.

Bcl-11b CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BCL11B expression without altering the underlying DNA sequence.

Bcl-11b CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BCL11B 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 BCL11B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Bcl-11b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BCL11B locus and enabling the study of Bcl-11b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Bcl-11b pathway restoration in tumor cells with silenced or reduced BCL11B expression.

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