ABCA5 CRISPR Activation Plasmid (h): sc-410568-ACT

ABCA5 encodes an ATP-binding cassette (ABC) transporter of the ABCA subfamily that localizes predominantly to intracellular membranes and contributes to ATP-dependent lipid translocation. By regulating intracellular cholesterol and sphingolipid handling, ABCA5 influences endolysosomal trafficking, membrane composition, and broader lipid homeostasis pathways. Altered ABCA5 expression has been linked to dysregulated sterol metabolism and cellular stress responses, and genetic variation has been investigated in the context of complex traits and lipid-associated pathophysiology. In cell models, ABCA5 provides a tractable node for dissecting how intracellular lipid transport interfaces with organelle function and metabolic signaling.

ABCA5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCA5 expression without altering the underlying DNA sequence.

ABCA5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCA5 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 ABCA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ABCA5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCA5 locus and enabling the study of ABCA5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ABCA5 pathway restoration in tumor cells with silenced or reduced ABCA5 expression.

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