@inbook{10fbe72f4ec941b5a093c77394079321,
title = "A recessive genetic screen for components of the RNA interference pathway in mouse embryonic stem cells",
abstract = "Several key components of the RNA interference (RNAi) pathway were identified in genetic screens performed in nonmammalian model organisms. To identify components of the mammalian RNAi pathway, we developed a recessive genetic screen in mouse embryonic stem (ES) cells. Recessive genetic screens are feasible in ES cells that are Bloom-syndrome protein (Blm-) deficient. Therefore, we constructed a reporter cell line in Blm-deficient ES cells to isolate RNAi mutants through a simple drug-selection scheme. This chapter describes how we used retroviral gene traps to mutagenize the reporter cell line and select for RNAi mutants. Putative RNAi mutants were confirmed using a separate functional assay. The location of the gene trap was then identified using molecular techniques such as Splinkerette PCR. Our screening strategy successfully isolated several mutant clones of Argonaute2, a vital component of the RNAi pathway.",
keywords = "Bloom-deficient mouse embryonic stem cells, RNA interference, recessive genetic screen, retroviral gene-trap mutagenesis, splinkerette PCR",
author = "Trombly, {Melanie I.} and Xiaozhong Wang",
year = "2010",
month = dec,
day = "1",
doi = "10.1007/978-1-60761-769-3_4",
language = "English (US)",
isbn = "9781607617686",
series = "Methods in Molecular Biology",
pages = "45--63",
editor = "Baohong Zhang and Edmund Stellwag",
booktitle = "RNAi and MicroRNA-Mediated Gene Regulation in Stem Cells",
}