The DNASE1L2 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human near-haploid HAP1 cell line. This model features targeted disruption of the DNASE1L2 gene, resulting in a loss-of-function background for investigating the role of this endonuclease in DNA degradation and keratinocyte terminal differentiation pathways.
The HAP1 parental cell line is a near-haploid chronic myeloid leukemia line with an adherent fibroblast-like morphology, originally established from the KBM-7 line. Its near-haploid karyotype facilitates straightforward genetic manipulation and eliminates confounding effects of multiple alleles, making it an ideal host for CRISPR-based knockout studies aimed at dissecting gene function and signaling networks.
DNASE1L2 encodes a DNase I-like endonuclease that degrades nuclear DNA during the terminal differentiation of keratinocytes, a prerequisite for cornified envelope assembly. The encoded enzyme operates downstream of the transcription factors TP53 and NOTCH1, integrating signals through effectors such as RBPJ and HES1. Its activity promotes the maturation of structural proteins including involucrin (IVL), loricrin (LOR), filaggrin (FLG), and transglutaminase 1 (TGM1), while also influencing the expression of CDKN1A, a p53 target. Consequently, DNASE1L2 is a pivotal node connecting DNA catabolism to epidermal barrier formation and nail development.
Although HAP1 cells are of leukemic origin and do not recapitulate keratinocyte differentiation, they provide a genetically tractable system to study the core molecular functions of DNASE1L2. The knockout model allows for the examination of DNASE1L2-dependent DNA degradation in a simplified context, the dissection of its regulation by the NOTCH1-p53 axis, and the identification of interacting partners that may link it to cornification. Defects in DNASE1L2 are associated with autosomal recessive nail dysplasia (NDNC4) and potential skin barrier abnormalities; therefore, this model supports research into the molecular mechanisms underlying these disorders.
Representative applications include functional genomics screens, TUNEL assays to quantify DNA fragmentation, immunofluorescence staining for differentiation markers such as KRT1 and KRT10, and western blotting or RT-qPCR to monitor pathway activity. These cells also enable the evaluation of modulators that influence the NOTCH1-DNASE1L2 signaling cascade and the search for compounds that can restore cornified envelope formation. For additional details or to discuss customized approaches, please contact Ascent Research.
