Lentiviral particles containing an inducible shRNA vector containing IQGAP1 targeting sequences (5’-GCCCACATTGTGCCTTTATTTCTC-3’, control 5’-CCTAAGGTTAAGTCGCCCTCGCTC-3’) or a scrambled control sequence cloned into the pLKO.Tet.On shRNA lentivector (Addgene, Cambridge, Mass) were generated as previously described.6 (link) Transduced PDAC cells were selected with 2 ug/mL puromycin and shRNAs were induced with 25 ng/mL (Capan-1) or 100 ng/mL (PANC-1) doxycycline (Sigma, St. Louis, Mo) for 3 or 7 days.
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Inducible IQGAP1 silencing in PDAC cells
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Corresponding Organization : The University of Texas at Austin
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Variable analysis
independent variables
- IQGAP1 targeting sequences (5'-GCCCACATTGTGCCTTTATTTCTC-3')
- Scrambled control sequence
- Proliferation of PDAC cells
- Puromycin concentration (2 ug/mL) for cell selection
- Doxycycline concentration (25 ng/mL for Capan-1, 100 ng/mL for PANC-1) for shRNA induction
- Induction duration (3 or 7 days)
- Positive control: IQGAP1 targeting shRNA
- Negative control: Scrambled control shRNA
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