PSENEN Knockout HaCaT Cell Line

The PSENEN Knockout HaCaT Cell Line is a CRISPR/Cas9-edited knockout model featuring targeted disruption of the human PSENEN gene in the HaCaT immortalized keratinocyte background. This cell line provides a stable loss-of-function system to analyze gamma-secretase complex activity. Supplied as a ready-to-use line, it eliminates the need for transient gene silencing, ensuring reproducible results in long-term studies.

HaCaT cells are a spontaneously immortalized human keratinocyte line derived from adult skin, maintaining the capacity for terminal differentiation when exposed to elevated calcium. This line models human epidermal biology, including proliferation, differentiation, and barrier formation. Its robust growth and genetic tractability make it widely used for gene-edited derivatives such as the PSENEN knockout.

PSENEN (PEN-2) is an essential subunit of the gamma-secretase complex, which also contains presenilin (PSEN1 or PSEN2), nicastrin (NCSTN), and anterior pharynx defective 1 (APH1A or APH1B). PSENEN facilitates presenilin endoproteolysis, a prerequisite for gamma-secretase maturation and catalytic activity. Once active, the complex mediates intramembrane proteolysis of type I transmembrane proteins: Notch receptors, amyloid precursor protein (APP), ErbB4, and E-cadherin. Cleavage of Notch generates the Notch intracellular domain (NICD), which transcriptionally activates targets such as Hes1 and Hey1. APP processing produces beta-amyloid peptides; ErbB4 cleavage releases its intracellular domain; E-cadherin processing yields fragments influencing adhesion. PSENEN knockout abolishes these events, thereby disrupting Notch, Wnt, Alzheimer’s, and ErbB signaling. Upstream regulators of PSENEN expression include SP1 and NF-kB, while Notch signaling provides feedback regulation.

In the HaCaT keratinocyte context, PSENEN knockout is particularly valuable for exploring gamma-secretase functions in epidermal homeostasis. HaCaT cells differentiate in response to calcium, a process governed in part by Notch-mediated transcriptional programs. Absence of PSENEN thus permits dissection of how gamma-secretase-dependent cleavage affects keratinocyte fate, intercellular adhesion via E-cadherin, and signaling cross-talk. This model is directly relevant to hidradenitis suppurativa, a chronic inflammatory skin disease linked to gamma-secretase mutations, as well as to Notch-related skin cancers and wound healing research.

Applications include high-throughput screening of gamma-secretase modulators using Notch reporter assays (e.g., CBF1-luciferase) or APP cleavage detection. Differentiation studies can be combined with RT-qPCR for Hes1 and Hey1, immunofluorescence for Notch1 and E-cadherin, and phospho-protein profiling. Transcriptomic (RNA-seq) and migration/scratch assays provide broader phenotypic readouts. This knockout line also serves as a non-neuronal platform for Alzheimer’s disease research exploring APP processing. For further details, contact Ascent Research.