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    Putting the Brakes on Metastatic Cancer

    June 21, 2018, Categories  Biology/Biotechnology

    Overview of the in vivo screen for genes required for productive motility. a HEp3 cells were transduced with a pooled whole human genome lentiviral shRNA library and injected intravenously into 100 ex ovo avian embryos. Compact metastatic colonies derived from single cancer cells were excised 6 days post injection, expanded and analyzed by Illumina deep sequencing. Colonies were re-injected individually to validate their phenotype and prioritized based on a composite compactness (C.I.) index. Selected screen hits that produced metastatic colonies that were significantly more compact than those produced by cancer cells transduced with scramble shRNA transduced cells were selected for further analysis. b Composite compactness index (C.I.) distribution of screen hits relative to positive (anti-CD151) and negative (scramble shRNA) controls. Screen hits that are significantly more compact than negative control are indicated in green. Clones containing a single shRNA species are in bold. For clones containing multiple shRNAs, the two most predominant shRNAs are shown. Statistical significance was determined using one-way ANOVA with Fisher’s LSD test (*p < 0.05, **p < 0.01, ***p < 0.001). c Table summarizing shRNA gene IDs from significantly compact clones containing single shRNA. Gene cards (http://www.genecards.org) and KEGG pathway (http://www.genome.jp) databases were used for gene function annotations

    Overview of the in vivo screen for genes required for productive motility. a HEp3 cells were transduced with a pooled whole human genome lentiviral shRNA library and injected intravenously into 100 ex ovo avian embryos. Compact metastatic colonies derived from single cancer cells were excised 6 days post injection, expanded and analyzed by Illumina deep sequencing. Colonies were re-injected individually to validate their phenotype and prioritized based on a composite compactness (C.I.) index. Selected screen hits that produced metastatic colonies that were significantly more compact than those produced by cancer cells transduced with scramble shRNA transduced cells were selected for further analysis. b Composite compactness index (C.I...

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