The HERPUD1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population with targeted disruption of the HERPUD1 gene, creating a loss-of-function model. The polyclonal format maintains genetic heterogeneity, offering a robust pool for functional studies without clonal isolation. This product is well-suited for investigating HERPUD1 function in endoplasmic reticulum (ER) stress signaling and ER-associated degradation (ERAD).
The host HAP1 cell line is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid genome ensures that knockout phenotypes are typically unambiguous, as there is only one allele to target. HAP1 cells are widely adopted in genetic screening and functional genomics applications due to their rapid growth, ease of CRISPR editing, and clean genetic background, making them ideal for mechanistic studies.
HERPUD1 is an ER-resident protein strongly induced by ER stress through the unfolded protein response (UPR), primarily downstream of transcription factors ATF6, XBP1, and ATF4. It functions as an adaptor that physically interacts with the HRD1-SEL1L ubiquitin ligase complex, promoting ubiquitination and retrotranslocation of misfolded ER proteins. The VCP/p97 ATPase then extracts substrates for proteasomal degradation, linking UPR to ERAD. HERPUD1 maintains ER homeostasis by facilitating clearance of aberrant proteins, and its activity involves cofactors such as Derlin-1, OS9, and XTP3-B.
In the HAP1 background, HERPUD1 knockout enables precise dissection of its contribution to ER proteostasis. Loss of HERPUD1 can be examined relative to the UPR branches??IRE1??/XBP1, PERK/eIF2??/ATF4, and ATF6??and their crosstalk with ERAD. This model helps elucidate how HERPUD1-dependent degradation of misfolded proteins influences cell fate under stress, providing insights into ER stress-related diseases like neurodegeneration and cancer.
Typical applications include western blot analysis of UPR markers (e.g., CHOP, GRP78), RT-qPCR for ER stress-responsive genes, proteasome activity assays, and cycloheximide chase experiments to monitor protein stability. The polyclonal population supports drug screening campaigns for proteostasis modulators and RNA-sequencing under ER stress to profile transcriptional changes. For further information, contact Ascent Research.
