FANCA CRISPR Activation Plasmid (m): sc-420284-ACT

Mouse Fanca encodes FANCA, a core component of the Fanconi anemia (FA) DNA repair pathway that safeguards genome stability during S phase. FANCA participates in the FA core complex that promotes monoubiquitination of FANCD2 and FANCI, coordinating recognition and repair of DNA interstrand crosslinks through crosstalk with homologous recombination and replication fork protection. Loss or dysfunction of FANCA compromises resolution of stalled replication intermediates, elevating chromosomal aberrations and hypersensitivity to crosslinking stress. Because the FA pathway is tightly linked to hematopoietic maintenance and tumor suppressive genome surveillance, Fanca is widely studied in models of inherited genome instability and cancer-relevant DNA damage responses.

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

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

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