Abstract
Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Cell Line
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DNA, Single-Stranded / genetics*
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Endonucleases / genetics
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Gene Deletion
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Gene Targeting / methods*
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Genetic Engineering / methods*
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Humans
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Mutagenesis, Insertional / methods
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Oligodeoxyribonucleotides / genetics
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Point Mutation / genetics
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Zinc Fingers / genetics
Substances
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DNA, Single-Stranded
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Oligodeoxyribonucleotides
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Endonucleases