Efficient, continuous mutagenesis in human cells using a pseudo-random DNA editor.

Haiqi Chen,Daniel Lesman,Fei Chen,Kettner Griswold,Allen Lin,Samuel Padula,Sophia Liu,Tongtong Zhao,Jamie L Marshall

doi:10.1038/s41587-019-0331-8

Efficient, continuous mutagenesis in human cells using a pseudo-random DNA editor.

Haiqi Chen, Daniel Lesman + Show 7 more

Open Access

https://doi.org/10.1038/s41587-019-0331-8

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Journal: Nature Biotechnology	Publication Date: Dec 16, 2019
Citations: 63

Affiliation: Broad Institute, Draper Laboratory, Massachusetts Institute of Technology, Harvard University, Center for Systems Biology, Ragon Institute of MGH, MIT and Harvard, Harvard–MIT Division of Health Sciences and Technology

#Mutagenesis In Human Cells #Multiple Cell Generations + Show 8 more

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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.

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