Titlebook: Chromosomal Mutagenesis; Gregory D. Davis,Kevin J. Kayser Book 2008 Humana Press 2008 DNA.Eukaryotic organisms.Evolution.Gene disruption.G

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Design and Testing of Zinc Finger Nucleases for Use in Mammalian Cells,cial proteins in which a zinc finger DNA-binding domain is fused to a nonspecific nuclease domain. This chapter describes how to identify potential targets for ZFN cutting, to make ZFNs to cut this target site, and how to test whether the newly designed ZFNs are active in a mammalian cell culture-based system.

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Highly Proficient Gene Targeting by Homologous Recombination in the Human Pre-B Cell Line Nalm-6,hnologies, gene knockout studies using human somatic cells will be of greater importance for analyzing the functions of human genes in greater detail. Although the frequency of gene targeting is typically very low in human cultured cells, we have recently shown that a human precursor B cell line, Na

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Design and Testing of Zinc Finger Nucleases for Use in Mammalian Cells, stem cells, and a few other specialized cell lines but has not been available to researchers in other systems, such as for mammalian somatic cell genetics. Recently, work has shown that the creation of a gene-specific DNA double-strand break can stimulate homologous recombination by several thousan

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Insertional Mutagenesis of the Mouse Germline With , Transposition, analyzed in an efficacious, controlled, and scalable manner. Thus, to facilitate analysis of gene function, new genetic tools and strategies are currently under development. Transposable elements, by virtue of their inherent ability to insert into DNA, can be developed into useful tools for chromos

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Conditional Gene Trapping Using the FLEx System,genes. It facilitates the recognition of sequence conservation, which helps to distinguish between important and not important and also coding from noncoding sequence. Nevertheless, approximately only 50% of all mouse genes have been entirely annotated to date. In the postgenomic era, large-scale pr

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Targeting Integration of the , Ty5 Retrotransposon, retroelement of specific chromatin states or DNA-bound protein complexes. A well-studied example of such a targeted retroelement is the . Ty5 retrotransposon, which integrates into heterochromatin at the telomeres and silent mating loci. Targeting is mediated by an interaction between Ty5 integrase