The EDEM1 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted gene disruption of the EDEM1 locus. This loss-of-function model leverages CRISPR/Cas9 to ablate EDEM1 expression, providing a versatile tool for studying glycoprotein quality control and ER-associated degradation (ERAD). The polyclonal format ensures a heterogeneous mixture of knockout alleles, enabling robust functional studies without single-cell cloning.
The HAP1 cell line is a near-haploid human fibroblast model derived from KBM-7 chronic myeloid leukemia (CML) cells. Its near-haploid karyotype simplifies genetic manipulation and knockout analysis, making it ideal for functional genomics. HAP1 cells retain features of adherent fibroblasts and are widely used in signal transduction and protein trafficking studies, providing a clean system for examining EDEM1-dependent pathways in a human context.
EDEM1 is an ERAD lectin that recognizes mannose-trimmed N-glycans on terminally misfolded glycoproteins, promoting their extraction from the calnexin cycle and targeting to the HRD1-SEL1L retrotranslocation complex for ubiquitination and proteasomal degradation. EDEM1 expression is upregulated by XBP1s and ATF6 under ER stress, linking UPR activation to enhanced protein clearance. It interacts with OS9, ERLEC1, derlin-1, and VCP/p97, ensuring efficient disposal of misfolded substrates and maintenance of ER homeostasis.
In the HAP1 background, EDEM1 knockout provides a unique model to dissect ERAD and UPR signaling without interference from redundant alleles, as the near-haploid nature ensures loss of function from a single allele disruption. This model is particularly relevant for studying glycoprotein quality control in fibroblasts derived from CML, linking ER stress to cancer biology. Researchers can examine how loss of EDEM1 sensitizes cells to proteotoxic stress, alters N-glycan processing, and affects stability of endogenous ERAD substrates.
This EDEM1 knockout polyclonal population supports diverse assays, including tunicamycin sensitivity tests, co-immunoprecipitation with SEL1L or HRD1, and RT-qPCR analysis of XBP1 splicing and UPR genes. The model is suitable for proteasome activity assays and ER immunofluorescence. Applications span from protein misfolding diseases and congenital disorders of glycosylation to cancer research and proteostasis drug screening. For more information, contact Ascent Research.
