USP20 Knockout Cell Lines

Gene: USP20

Official Full Name: ubiquitin specific peptidase 20provided by HGNC

Gene Summary: This gene encodes a ubiquitin specific processing protease that was first identified as a substrate of the VHL (von Hippel-Lindau disease) protein E3 ubiquitin ligase complex. In addition to being ubiquitinated by the VHL-E3 ligase complex, this enzyme deubiquitinates hypoxia-inducible factor (HIF)-1 alpha and thereby causes increased expression of HIF-1alpha targeted genes which play a role in angiogenesis, glucose metabolism, cell proliferation and metastasis. The enzyme encoded by this gene also regulates G-protein coupled receptor signaling by mediating the deubiquitination of beta-2 adrenergic receptor (ADRB2). This enzyme is a ubiquitously expressed thiolester hydrolase. Alternative splicing results in multiple transcript variants encoding the same protein. [provided by RefSeq, Jan 2013]

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Products Background Case study

Products

Catalog Number	Product Name	Species	Gene	Passage ratio	Mycoplasma testing	Price
KO00005	USP20 Knockout cell line （THP-1）	Human	USP20	1：2-1：4	Negative	Online Inquiry
KO05798	USP20 Knockout cell line (HeLa)	Human	USP20	1:3~1:6	Negative	Online Inquiry
KO05799	USP20 Knockout cell line (HCT 116)	Human	USP20	1:2~1:4	Negative	Online Inquiry
KO05800	USP20 Knockout cell line (HEK293)	Human	USP20	1:3~1:6	Negative	Online Inquiry
KO05801	USP20 Knockout cell line (A549)	Human	USP20	1:3~1:4	Negative	Online Inquiry

Background

USP20 Gene Knockout Cell Lines consist of genetically modified cell lines in which the USP20 gene, a pivotal component in cellular regulatory pathways, has been systematically inactivated. This innovative product facilitates the study of the biological role of USP20, which encodes a deubiquitinating enzyme involved in modulating various cellular processes such as protein stability, intracellular signaling, and immune responses.

Key functions of USP20 Gene Knockout Cell Lines revolve around their ability to elucidate the effects of USP20 loss on cellular dynamics and pathways, providing researchers with a powerful tool for investigating the enzyme's influence on the ubiquitin-proteasome system. By leveraging these cell lines, scientists can conduct a myriad of experiments that explore the functional implications of USP20 knockout, advancing our understanding of diseases associated with dysregulated ubiquitin pathways and potentially facilitating drug discovery and development.

The scientific importance of these cell lines is particularly pronounced in translational research settings, where the proper functioning of the ubiquitin-proteasome system is critical in the context of cancer, neurodegenerative conditions, and immunological disorders. Hence, applications extend from basic research to potential therapeutic interventions, making USP20 Gene Knockout Cell Lines invaluable for both researchers and clinicians aiming to decode the complexities of protein regulation.

Compared to alternative cell models, USP20 Gene Knockout Cell Lines are designed with high specificity and reproducibility, minimizing off-target effects and maximizing experimental integrity. Their optimized use in high-throughput screening platforms sets them apart, enabling researchers to efficiently evaluate pharmacological candidates in a genetically controlled environment.

The value of USP20 Gene Knockout Cell Lines lies in their unique ability to provide robust data and insights into the role of deubiquitinating enzymes, which are often overlooked in traditional studies. By choosing our product, researchers and clinicians are equipped with a precise tool that enhances experimental outcomes and drives innovation in the understanding of complex cellular processes.

With our extensive expertise in cellular biology and genetic engineering, we are committed to providing high-quality biological products that empower researchers and contribute to advancements in life sciences.

Case study

USP20 Gene Knockout Cell Lines: Deciphering the Role of USP20 in Reticulophagy and ER Homeostasis

Research Focus

Investigate the function of USP20 (ubiquitin specific peptidase 20) in reticulophagy (ER-phagy), a selective autophagic process that degrades impaired endoplasmic reticulum (ER) subdomains, and its mechanism of action in maintaining ER homeostasis.

Background

1. Reticulophagy and ER Homeostasis

The ER is essential for protein synthesis, lipid metabolism, and calcium storage. Reticulophagy selectively removes damaged ER regions to maintain cellular balance. Dysregulation of this process is linked to neurodegenerative diseases and metabolic disorders .

2. Role of Ubiquitination in Autophagy

Ubiquitination and deubiquitination (DUB) enzymes regulate autophagic pathways. However, their specific roles in reticulophagy remain unclear. USP20, a DUB, was identified as a potential regulator due to its ability to cleave ubiquitin chains and stabilize target proteins .

3. Gaps in Knowledge

How does USP20 influence reticulophagy?

What is the mechanism by which USP20 regulates ER-resident proteins?

Can USP20 knockout (KO) cell lines serve as a model to study ER-related diseases?

Solution

1. Generation of USP20 KO Cell Lines

Method: CRISPR-Cas9 genome editing was used to disrupt the USP20 gene in HeLa and U-2 OS cells. Guide RNAs targeted the USP20 coding sequence, and KO was confirmed by western blotting and immunofluorescence .

Validation: USP20 KO cells showed reduced LC3B-II levels and impaired autophagic flux, as evidenced by decreased autolysosome formation (red puncta) and accumulated autophagosomes (yellow puncta) under starvation (EBSS) conditions .

2. Functional Assays in USP20 KO Cells

Reticulophagy Impairment: KO cells exhibited decreased cleavage of GFP-SEC61B (an ER marker) and reduced red puncta formation in the ssRFP-GFP-KDEL assay, indicating defective reticulophagy .

RETREG1/Ubiquitination Analysis: USP20 KO cells showed increased K48- and K63-linked ubiquitination of RETREG1 (a reticulophagy receptor), leading to RETREG1 degradation via the proteasome pathway .

Subcellular Localization: USP20 KO disrupted the interaction between RETREG1 and VAPs (VAMP-associated proteins), impairing ER recruitment of autophagy proteins like WIPI2 .

3. Rescue Experiments

Reintroduction of wild-type USP20, but not catalytically inactive (USP20C154S,H643Q) or FFAT motif-deleted mutants, restored reticulophagy in KO cells, confirming USP20’s role in deubiquitinating RETREG1 .

Conclusion

1. Mechanistic Insights

USP20 regulates reticulophagy by:

Deubiquitinating K48- and K63-linked chains on RETREG1, stabilizing the receptor .

Interacting with VAPs via FFAT motifs to localize to ER subdomains, promoting recruitment of WIPI2 and autophagy initiation .

2. Translational Significance

Disease Modeling: USP20 KO cells recapitulate ER dysfunction seen in neurodegenerative diseases (e.g., mutations in reticulophagy components) .

Drug Discovery: USP20 may serve as a target for diseases involving ER stress, such as Alzheimer’s or metabolic disorders .

3. Product Utility

USP20 KO cell lines provide a robust model for:

Studying reticulophagy pathways and ER homeostasis.

Identifying USP20-interacting proteins and downstream effectors.

Screening compounds that modulate USP20 activity or RETREG1 ubiquitination.

USP20 Gene Knockout Cell Lines

Please note that all services are for research use only. Not intended for any clinical use.

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