Abstract
Here we describe TRACE (T7 polymerase-driven continuous editing), a method that enables continuous, targeted mutagenesis in human cells using a cytidine deaminase fused to T7 RNA polymerase. TRACE induces high rates of mutagenesis over multiple cell generations in genes under the control of a T7 promoter integrated in the genome. We used TRACE in a MEK1 inhibitor-resistance screen, and identified functionally correlated mutations.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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DNA / genetics*
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DNA-Directed RNA Polymerases / metabolism
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Gene Editing / methods*
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Genetic Loci
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HEK293 Cells
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Humans
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Mutagenesis / genetics*
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Mutation / genetics
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Promoter Regions, Genetic
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Viral Proteins / metabolism
Substances
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Viral Proteins
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DNA
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bacteriophage T7 RNA polymerase
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DNA-Directed RNA Polymerases