The JOSD1 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population in which the JOSD1 gene has been disrupted. This mixture of knockout cells carries a variety of gene-inactivating mutations, generating a loss-of-function model that avoids clonal artifacts. JOSD1 encodes a deubiquitinase that cleaves ubiquitin chains, modulating protein degradation and signaling. This product enables detailed functional investigations of JOSD1 in a cancer cell background without the need for single-cell cloning.
The HCT 116 host cell line is a human colorectal carcinoma epithelial model harboring a KRAS G13D mutation and high microsatellite instability (MSI-high) due to MLH1 deficiency. These oncogenic and mutator phenotypes establish it as a widely used system for colorectal cancer research. The cell line??s adherent growth and well-characterized genetics support robust experimental manipulation. JOSD1 knockout in this background links deubiquitinase function to established oncogenic pathways and DNA repair defects.
JOSD1 functions as a deubiquitinase that hydrolyzes ubiquitin chains, thereby regulating protein turnover and non-proteolytic signaling events. It interacts with ubiquitin and proteasome subunits to remove ubiquitin moieties from substrate proteins, potentially reversing degradation signals or altering protein interactions. This activity is integral to the ubiquitin-proteasome system and influences endocytic trafficking and autophagy. While upstream regulators remain poorly defined, JOSD1 likely responds to cellular stress cues. Downstream targets include ubiquitinated transmembrane receptors and signaling proteins, whose stability and activity it governs. Together with E3 ubiquitin ligases and the 26S proteasome, JOSD1 maintains the dynamic equilibrium of ubiquitin conjugation. Disruption of JOSD1 perturbs this balance, offering a means to dissect its substrate-specific roles.
In HCT 116 colorectal cancer cells, JOSD1 knockout may profoundly alter oncogenic signaling and cellular responses due to the host line??s genetic vulnerabilities. The KRAS mutation drives proliferation, while MSI-high status indicates defective DNA mismatch repair; aberrant deubiquitination could impact both growth control and genomic maintenance. Loss of JOSD1 activity likely leads to accumulation of ubiquitinated proteins, affecting signal transduction, endosomal sorting, and autophagy. This model allows dissection of whether JOSD1 supports or suppresses tumorigenic phenotypes, thereby clarifying its functional role in colorectal cancer pathogenesis.
This polyclonal knockout product is suitable for a variety of experimental approaches. Researchers can confirm JOSD1 loss and monitor global ubiquitin profiles via western blotting, and assess protein half-life using cycloheximide chase assays. Co-immunoprecipitation can reveal altered ubiquitin-dependent protein complexes, while immunofluorescence detects changes in subcellular localization. Drug sensitivity panels with oxaliplatin or 5-fluorouracil, coupled with caspase-3/7 apoptosis assays, can evaluate therapeutic vulnerabilities. Migration and invasion assays further probe metastatic potential. As a pooled knockout resource, these cells facilitate robust functional genomics studies. For further product information, please contact Ascent Research.
