The IRAK2 Knockout HaCaT Polyclonal Cells represent a CRISPR/Cas9-edited human polyclonal knockout population targeting the IRAK2 gene in the HaCaT keratinocyte cell line. The pooled editing strategy yields a heterogeneous mix of cells, each carrying distinct gene-disrupting modifications, providing a loss-of-function model that avoids clonal artifact.
HaCaT is a spontaneously immortalized, non-tumorigenic human keratinocyte line extensively used for studies of epidermal differentiation, innate immunity, and inflammatory skin diseases. These cells retain key keratinocyte features, including the ability to secrete cytokines and chemokines upon immune challenge, making them a valuable host for dissecting signaling pathways relevant to skin homeostasis.
IRAK2 functions as a signaling adaptor downstream of IL-1R and TLRs. Upon receptor activation by ligands such as IL-1?? or LPS, IRAK2 is recruited to MyD88 and forms a complex with IRAK4 and TRAF6, enabling activation of TAK1 and the IKK complex. This leads to NF-??B and MAP kinase (JNK, p38) activation, which drive expression of pro-inflammatory cytokines including IL-6, IL-8, and TNF-??, as well as chemokines.
In keratinocytes, IRAK2-mediated signaling is pivotal for innate immune responses. Keratinocytes secrete cytokines and antimicrobial peptides upon IL-1/TLR stimulation, and IRAK2 is a key node in this process. Dysregulation of this pathway contributes to inflammatory skin pathologies such as psoriasis and atopic dermatitis. The IRAK2 Knockout HaCaT Polyclonal Cells thus offer a physiologically relevant tool to study how IRAK2 loss affects keratinocyte immune function and barrier integrity.
This polyclonal knockout population is suited for a range of assays, including immunoblotting for phospho-I??B?? and phospho-p65, RT-qPCR analysis of IL-6 and IL-8 transcripts, and ELISA quantification of secreted cytokines. It can be employed to examine IRAK2 interactions with MyD88 and TRAF6 by co-immunoprecipitation, to assess NF-??B nuclear translocation via immunofluorescence, and to evaluate keratinocyte migration or barrier function. The model supports screening of pathway modulators and anti-inflammatory compounds in a keratinocyte context. For further details, please contact Ascent Research.
