Bcl-11a CRISPR Activation Plasmid (h): sc-403612-ACT

BCL11A encodes the transcription factor Bcl-11a, a zinc finger DNA-binding protein that regulates lineage specification and differentiation programs, particularly within hematopoietic and immune compartments. It functions in chromatin-associated transcriptional control, integrating with coregulatory complexes to shape gene expression networks involved in cell fate decisions, proliferation, and developmental timing. BCL11A is closely linked to erythroid globin regulation and broader epigenetic control of hematopoietic maturation, making it a key node in pathways governing hemoglobin switching and lymphoid development. Dysregulated BCL11A activity has been associated with hematologic disease biology and altered differentiation states, supporting its use as a mechanistic target in functional genomics studies.

Bcl-11a CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BCL11A expression without altering the underlying DNA sequence.

Bcl-11a CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BCL11A 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 BCL11A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Bcl-11a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BCL11A locus and enabling the study of Bcl-11a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Bcl-11a pathway restoration in tumor cells with silenced or reduced BCL11A expression.

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