The ABCA12 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human ABCA12 gene in HEK293T cells. This loss-of-function model disrupts the ATP-binding cassette transporter responsible for loading glucosylceramides into lamellar granules, a critical step in epithelial lipid barrier formation. As a polyclonal pool, the cells maintain genetic diversity while ensuring robust gene ablation across the population.
HEK293T is an immortalized human embryonic kidney epithelial cell line transformed with SV40 large T antigen, offering high transfection efficiency and a well-characterized genome. Although not of keratinocyte origin, its epithelial background supports studies of membrane transport and lipid metabolism, making it suitable for dissecting ABCA12-dependent trafficking pathways in a tractable in vitro system.
ABCA12 encodes a glucosylceramide transporter essential for lamellar body loading and subsequent epidermal lipid barrier formation. It is transcriptionally regulated by PPAR??, RAR, and RXR, and functionally interacts with TGM1, LIPH, ALOX12B, ALOXE3, and ceramide synthases. ABCA12 activity promotes lamellar body secretion, TGM1-mediated crosslinking of involucrin and loricrin, and corneocyte lipid envelope assembly. Knockout of ABCA12 ablates glucosylceramide trafficking, impairing the permeability barrier and mimicking the molecular defects of harlequin ichthyosis.
In HEK293T cells, ABCA12 knockout provides a reductionist model to study lipid transport without the complexity of terminal keratinocyte differentiation. This system facilitates reconstitution studies with upstream regulators or downstream effectors to dissect epistatic relationships and screen for modulators of glucosylceramide trafficking. It is particularly suited for evaluating PPAR??/RXR agonists or ceramide synthesis inhibitors that may restore lipid delivery, and for co-expression assays with TGM1 or ALOX enzymes to map the epidermal lipid assembly pathway.
Typical applications include Western blotting for ABCA12, immunostaining of lamellar body markers, lipidomics profiling of ceramide species, and RT-qPCR quantification of differentiation genes (involucrin, loricrin). Reporter assays for PPAR and RXR activity enable dissection of upstream signaling cascades. These cells are valuable for screening therapeutics for ichthyosis and other skin barrier disorders, and for fundamental research into lipid transporter function in epithelial biology. For further information, please contact Ascent Research.
