The EIF2AK3 Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human DLD-1 colorectal adenocarcinoma cell line. This product utilizes targeted gene disruption to eliminate PERK kinase expression, creating a heterogeneous loss-of-function model suitable for population-based functional analyses. The polyclonal format captures a range of editing events, facilitating robust phenotypic assessment of the UPR without clonal isolation.
The DLD-1 host cell line is an epithelial colorectal adenocarcinoma model carrying well-characterized mutations in APC, KRAS, and TP53. These alterations result in constitutive WNT pathway activation, persistent RAS?CMAPK signaling, and compromised p53-mediated cell cycle arrest and apoptosis. This genetic profile mirrors aggressive colorectal tumors, providing a disease-relevant system to study stress adaptation and oncogenic signaling crosstalk.
EIF2AK3 encodes PERK, an ER-transmembrane kinase central to the unfolded protein response. Under ER stress, BiP/GRP78 dissociation triggers PERK autophosphorylation and activation, leading to phosphorylation of eIF2?? at Ser51. This modification transiently inhibits global protein synthesis while selectively enhancing ATF4 translation. ATF4 transactivates genes including the pro-apoptotic transcription factor CHOP (DDIT3) and the negative feedback regulator GADD34. PERK also directly phosphorylates NRF2, promoting its dissociation from KEAP1 and activating antioxidant target genes. PERK signaling intersects with the IRE1 and ATF6 UPR branches and involves interacting partners such as TRB3 and DNAJC3. Upstream stressors include tunicamycin, thapsigargin, hypoxia, nutrient deprivation, and reactive oxygen species, collectively dictating cell fate decisions between survival and apoptosis.
In the DLD-1 background, PERK disruption is particularly informative given the elevated ER stress imposed by oncogenic KRAS-driven protein synthesis and loss of p53-mediated apoptotic control. The polyclonal knockout pool allows evaluation of PERK dependency in colorectal cancer cell proliferation, migration, and resistance to ER stress-inducing chemotherapeutics. This model avoids clonal selection biases, enabling unbiased assessment of PERK??s contribution to tumor cell fitness under conditions such as hypoxia or nutrient scarcity.
Typical applications encompass mechanistic UPR studies, drug sensitivity profiling, and exploration of PERK-mediated signaling in colorectal cancer. Representative assays include Western blot for phospho-eIF2?? (Ser51) and ATF4, RT-qPCR for CHOP and GADD34, ATF4 luciferase reporter, Annexin V flow cytometry, co-immunoprecipitation of PERK-BiP, and immunofluorescence for PERK localization. These cells also enable RNA-seq-based transcriptome analysis and high-throughput screening of UPR modulators. For further technical details, please contact Ascent Research.
