@inbook{0c5c1aa2a82444e9a49245c1ab6e7383,
title = "Ex Vivo Transposon-Mediated Genetic Screens for Cancer Gene Discovery",
abstract = "Transposon mutagenesis has emerged as a powerful methodology for functionally annotating cancer genomes. Although in vivo transposon-mediated forward genetic screens have proven to be valuable for cancer gene identification, they are also time consuming and resource intensive. To facilitate the rapid and cost-effective identification of genes that regulate tumor-promoting pathways, we developed a complementary ex vivo transposon mutagenesis approach wherein human or mouse cells growing in culture are mutagenized and screened for the acquisition of specific phenotypes in vitro or in vivo, such as growth factor independence or tumor-forming ability. This approach allows discovery of both gain- and loss-of-function mutations in the same screen. Transposon insertions sites are recovered by high-throughput sequencing. We recently applied this system to comprehensively identify and validate genes that promote growth factor independence and transformation of murine Ba/F3 cells. Here we describe a method for performing ex vivo Sleeping Beauty-mediated mutagenesis screens in these cells, which may be adapted for the acquisition of many different phenotypes in distinct cell types.",
keywords = "Cell-based screens, DNA transposons, Ex vivo forward genetic screening, Functional annotation, Growth-factor independence, Sleeping Beauty, Transposon mutagenesis",
author = "O'Donnell-Mendell, {Kathryn A} and Yabin Guo and Shruthy Suresh and Updegraff, {Barrett L.} and Xiaorong Zhou",
note = "Funding Information: This work was supported by grants R01CA207763 from the National Cancer Institute (to K.A.O.), NIH Awards (T32GM008203 to B.L.U. and T32GM10977601 to S.S.), I-1881 from The Welch Foundation (to K.A.O.), SKF-15-067 from the Sidney Kimmel Foundation (to K.A.O.), 2015-03 from The LUNGevity Foundation (to K.A.O.), a HHMI Med into Grad Grant (56006776 to S.S.), and the Cancer Prevention Research Institute of Texas (CPRIT, RP150676 to K.A.O, RP140110 to B. L.U and S.S., and RP160157 to S.S.). We thank Jose Cabrera for assistance with the figures.",
year = "2019",
doi = "10.1007/978-1-4939-8967-6_12",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "145--157",
booktitle = "Methods in Molecular Biology",
}