Advisor(s)

Dr. Maurizio Bocchetta, Loyola Medical Center

Location

Room Academic Pit

Start Date

26-4-2019 2:10 PM

End Date

26-4-2019 2:35 PM

Abstract

Deubiquitinases (DUBs) are crucial determinants of protein stability, localization, complex formation and activity within cells (1). The OTUD6B DUB has been previously linked to cell growth and proliferation (2). More specifically, the isoform OTUD6B-2 may promote protein synthesis and DNA synthesis through deubiquitination of c-Myc (2). We established OTUD6B-knockout A549 and H1299 non-small cell lung cancer (NSCLC) cell lines to determine how OTUD6B contributes to the malignant potential of cancer. To delete OTUD6B expression, we utilized clustered regularly interspaced short palindromic repeats (CRISPR) technology, using short RNA guides to direct an endonuclease (Cas9) to specific sites on genomic DNA (3). Our approach involved double nicking/homologous recombination insertion (4) of an antibiotic resistance cassette into the OTUD6B exon IV.

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Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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Apr 26th, 2:10 PM Apr 26th, 2:35 PM

CRISPR knockout of the DUB OTUD6B in lung cancer cells

Room Academic Pit

Deubiquitinases (DUBs) are crucial determinants of protein stability, localization, complex formation and activity within cells (1). The OTUD6B DUB has been previously linked to cell growth and proliferation (2). More specifically, the isoform OTUD6B-2 may promote protein synthesis and DNA synthesis through deubiquitination of c-Myc (2). We established OTUD6B-knockout A549 and H1299 non-small cell lung cancer (NSCLC) cell lines to determine how OTUD6B contributes to the malignant potential of cancer. To delete OTUD6B expression, we utilized clustered regularly interspaced short palindromic repeats (CRISPR) technology, using short RNA guides to direct an endonuclease (Cas9) to specific sites on genomic DNA (3). Our approach involved double nicking/homologous recombination insertion (4) of an antibiotic resistance cassette into the OTUD6B exon IV.

 

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