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Toward Multiplex Genome Engineering in Mammalian Cells

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2013-10-18

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Rios Villanueva, Xavier. 2013. Toward Multiplex Genome Engineering in Mammalian Cells. Doctoral dissertation, Harvard University.

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Abstract

Given the explosion in human genetic data, new high-throughput genetic methods are necessary for studying variants and elucidating their role in human disease. In Chapter I, I will expand on this concept and describe current methods for genetically modifying human cells. In E. coli, Multiplex Automatable Genome Engineering (MAGE) is a powerful tool that enables the targeting of multiple genomic loci simultaneously with synthetic oligos that are recombined at high frequencies in an optimized strain. MAGE as a method has two components: organism-specific optimization of oligo recombination parameters and a protein capable of increasing recombination frequencies.

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Genetics, Biophysics, Genome Engineering, recombination, Synthetic Biology

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