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Generation of SMURF2 knockout human cells using the CRISPR/Cas9 system
Dhanoop Manikoth Ayyathan1, Nataša Ilić1, Hava Gil-Henn2
1Laboratory of Molecular and Cellular Cancer Biology, Faculty of Medicine in the Galilee, Bar-Ilan University, Safed, Israel.
Abstract:
The HECT domain E3 ubiquitin ligase SMURF2 regulates stability of several key protein targets involved in tumorigenesis, cell proliferation, migration, differentiation, and senescence. While altered levels and aberrant cellular distribution of SMURF2 were reported in different types of cancer, its role in tumorigenesis is far from understood. To elucidate the role of SMURF2 in cancer, appropriate human cancer cell models are needed. Here, we describe approaches that can be used to generate human normal and cancer cell strains knocked-out for SMURF2 using the clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) gene-editing technology.
Insights
Researchers developed methods to create cancer cell models lacking SMURF2, a protein linked to tumor growth. These models are crucial for understanding SMURF2's role in cancer development and progression.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- SMURF2 (HECT domain E3 ubiquitin ligase) influences protein targets critical for cell processes like proliferation and migration.
- Altered SMURF2 levels are observed in various cancers, but its precise role in tumorigenesis remains unclear.
Purpose of the Study:
- To investigate the function of SMURF2 in cancer.
- To establish human normal and cancer cell models with SMURF2 knocked out.
Main Methods:
- Utilized CRISPR/Cas9 gene-editing technology.
- Generated human normal and cancer cell strains lacking SMURF2.
Main Results:
- Successfully created human cell models with SMURF2 gene knockout.
- These models provide a platform for studying SMURF2's function in cancer.
Conclusions:
- The developed CRISPR/Cas9-based approach enables the generation of essential cellular tools for cancer research.
- These SMURF2-knockout models will aid in elucidating SMURF2's involvement in tumorigenesis.
