CTRL and SGEF KD MDCK cell lines were established in our previous publication (Awadia et al., 2019 (link)). Constructs to KD Slug in SGEF KD were cloned into pLKO.1-Hygro (Addgene 24150). To generate Scribble and Dlg1 CRISPR/Cas9 KO cell lines we used pLentiCRISPR v2-Blast (Addgene 83480). DH-PH and VSV-G-DH-PH Rescue constructs were cloned using gateway recombination (Thermo Fisher) into pLenti CMV Hygro DEST (Addgene 17454). We generated stable cells lines using these constructs via lentiviral delivery and antibiotic selection. For KO cell lines single cell clones were identified and validated. Details for the constructs used in this study, including shRNA and gRNA targeting sequences utilized are listed in Supp. Table 1
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Establishing Stable Cell Lines for Molecular Studies
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Other organizations : University of Toledo
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Variable analysis
independent variables
- Scribble CRISPR/Cas9 KO
- Dlg1 CRISPR/Cas9 KO
- Overexpression of DH-PH and VSV-G-DH-PH
- Not explicitly mentioned
- CTRL and SGEF KD MDCK cell lines
- Positive control: CTRL cell line
- Negative control: SGEF KD cell line
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