Efficient, continuous mutagenesis in human cells using a pseudo-random DNA editor
2019
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. Efficient diversification of DNA sequences is enabled by a T7 RNA polymerase fused to a cytidine deaminase.
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