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Generation of IL17RB Knockout Cell Lines Using CRISPR/Cas9-Based Genome Editing
Olivia Hu1, Alessandro Provvido1, Yan Zhu2
1Department of Biological Sciences, St. John's University, Queens, NY, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 16, 2020
Summary
We developed a CRISPR/Cas9 method to create knockout cell lines for interleukin-17 receptor B (IL17RB). This efficient gene editing technique enables the study of IL17RB
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Background:
- CRISPR/Cas9 is a powerful tool for genetic modification.
- Interleukin-17 receptor B (IL17RB) is overexpressed in various cancers and binds IL17B and IL17E.
- Establishing knockout cell lines is crucial for studying gene function.
Purpose of the Study:
- To present a methodological approach for generating interleukin-17 receptor B (IL17RB) knockout cell lines.
- To demonstrate the utility of CRISPR/Cas9-mediated genomic deletion for creating specific gene knockouts.
- To provide a framework for generating other mammalian gene knockout cell lines.
Main Methods:
- CRISPR design and cloning.
- Delivery of CRISPR constructs into cells.
- Verification of gene deletion using PCR and genomic DNA extraction.
Main Results:
- Successfully generated IL17RB knockout cell lines using CRISPR/Cas9.
- Validated the efficiency of the genomic deletion method.
- Confirmed the absence of IL17RB gene expression.
Conclusions:
- The presented CRISPR/Cas9 method is effective for creating IL17RB knockout cell lines.
- This approach offers a reliable strategy for generating gene knockouts in mammalian cells.
- The generated cell lines can facilitate research into the role of IL17RB in cancer.
