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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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.