FOXD2 CRISPR Activation Plasmid (h): sc-410785-ACT

FOXD2 encodes a forkhead box (FOX) family transcription factor that binds sequence-specific DNA motifs to regulate gene expression programs involved in cellular identity and developmental patterning. As a nuclear regulator, FOXD2 contributes to transcriptional networks controlling lineage specification, differentiation, and tissue morphogenesis through coordinated modulation of downstream target genes. Dysregulated FOX-family transcriptional control is frequently linked to aberrant proliferation and altered differentiation states, making FOXD2 relevant to studies of developmental disorders and cancer-associated gene regulatory circuitry. Human FOXD2 is therefore useful for interrogating how transcription factor dosage reshapes chromatin-dependent transcriptional outputs in model systems.

FOXD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXD2 expression without altering the underlying DNA sequence.

FOXD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXD2 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 FOXD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXD2 locus and enabling the study of FOXD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXD2 pathway restoration in tumor cells with silenced or reduced FOXD2 expression.

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