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- W2052334244 abstract "Genome editing followed by sequencing has now been used to engineer and analyse every variation of several stretches of human DNA in living cells, providing insight into the function of each constituent nucleotide molecule. See Letter p.120 There is a large demand in the field of genomics for techniques that can rapidly and cost-effectively determine the functional consequences of mutations. This paper describes a way of performing saturation mutagenesis of genomic regions — aiming to generate all possible mutations — whilst maintaining the native endogenous chromosomal context. The method uses CRISPR/Cas9 RNA-guided cleavage and multiplex homology-directed repair; its utility is demonstrated within exon 18 of BRCA1 by replacement of a six base-pair genomic region with all possible hexamers and by creating the same whole exon with all possible single nucleotide variants. Saturation editing was also performed for a well-conserved coding region of an essential gene, DBR1. The approach promises to facilitate high-resolution functional dissection of cis-regulatory elements and trans-acting factors, and the interpretation of variants of uncertain significance reported by clinical sequencing." @default.
- W2052334244 created "2016-06-24" @default.
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- W2052334244 date "2014-08-20" @default.
- W2052334244 modified "2023-10-04" @default.
- W2052334244 title "Edit the genome to understand it" @default.
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- W2052334244 doi "https://doi.org/10.1038/nature13659" @default.
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