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Nat Biotechnol ; 38(2): 165-168, 2020 02.
Article in English | MEDLINE | ID: mdl-31844291

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.


Subject(s)
DNA/genetics , Gene Editing/methods , Mutagenesis/genetics , DNA-Directed RNA Polymerases/metabolism , Genetic Loci , HEK293 Cells , Humans , Mutation/genetics , Promoter Regions, Genetic , Viral Proteins/metabolism
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