Aldehyde dehydrogenase 1-A2/ALDH1A2 CRISPR Activation Plasmid (h): sc-401340-ACT

Human ALDH1A2 encodes aldehyde dehydrogenase 1-A2, a cytosolic retinaldehyde dehydrogenase that catalyzes oxidation of retinaldehyde to retinoic acid, a key morphogen controlling transcriptional programs via retinoic acid receptors. Through regulation of retinoid metabolism and RA-dependent gene expression, ALDH1A2 influences embryonic patterning, epithelial differentiation, and tissue homeostasis, and intersects with developmental signaling networks such as HOX gene regulation. Altered ALDH1A2 activity or expression can perturb retinoic acid gradients and has been associated with congenital malformations and context-dependent changes in differentiation state relevant to cancer and fibrotic disease biology. Because retinoic acid signaling integrates metabolic state with transcriptional control, ALDH1A2 serves as a useful node for dissecting pathway crosstalk and cell fate decisions.

Aldehyde dehydrogenase 1-A2/ALDH1A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ALDH1A2 expression without altering the underlying DNA sequence.

Aldehyde dehydrogenase 1-A2/ALDH1A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ALDH1A2 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 ALDH1A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Aldehyde dehydrogenase 1-A2/ALDH1A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ALDH1A2 locus and enabling the study of Aldehyde dehydrogenase 1-A2/ALDH1A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Aldehyde dehydrogenase 1-A2/ALDH1A2 pathway restoration in tumor cells with silenced or reduced ALDH1A2 expression.

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