The ARL6IP1 Knockout HEK293T Polyclonal Cells from Ascent Research are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ARL6IP1 gene. This loss-of-function model provides a versatile tool for investigating endoplasmic reticulum (ER) dynamics, apoptosis, and intracellular trafficking. The polyclonal nature supports pooled functional analyses while maintaining robust target-gene disruption.
These cells are derived from HEK293T, a human embryonic kidney epithelial line transformed with adenovirus 5 DNA and stably expressing SV40 large T antigen. HEK293T permits high-efficiency transient transfection and episomal plasmid replication, making it a preferred host for recombinant protein production, viral vector generation, and gene-delivery studies.
ARL6IP1 encodes an ER-shaping protein that promotes ER tubulation and functions as an anti-apoptotic factor by preserving mitochondrial membrane potential and preventing cytochrome c release. It interacts with ARL6, atlastins, reticulons, and BCL-2 family members, and is regulated by ER stress sensors (PERK, IRE1) and apoptotic stimuli. Downstream, ARL6IP1 inhibits caspase-9 and caspase-3 activation, placing it at a critical node linking ER morphology to apoptosis.
Disruption of ARL6IP1 in HEK293T cells creates a model to study ER structural dynamics and cytoprotective signaling in an epithelial background. The polyclonal knockout population allows investigation of ARL6IP1-dependent pathways relevant to hereditary spastic paraplegia type 61 (SPG61), cancer cell survival, and viral entry, all of which involve ER remodeling and apoptosis modulation.
Applications include ER morphology analysis by immunofluorescence (calnexin/KDEL), apoptosis assays (caspase-3/7, Annexin V/PI), cytochrome c release and mitochondrial membrane potential measurements (JC-1/TMRE). ER stress can be induced with tunicamycin or thapsigargin, and ARL6 interactions examined by co-immunoprecipitation. Viral entry studies employ HIV-1 pseudoviruses, complemented by RT-qPCR, Western blot, and cell viability assays (MTT/XTT). For more information, contact Ascent Research.
