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

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

doi:10.1038/s41587-019-0331-8

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

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

Research output: Contribution to journal › Article › peer-review

47 Scopus citations

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.

Original language	English (US)
Pages (from-to)	165-168
Number of pages	4
Journal	Nature biotechnology
Volume	38
Issue number	2
DOIs	https://doi.org/10.1038/s41587-019-0331-8
State	Published - Feb 1 2020
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology
Molecular Medicine
Biomedical Engineering

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10.1038/s41587-019-0331-8

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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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AU - Chen, Haiqi

AU - Liu, Sophia

AU - Padula, Samuel

AU - Lesman, Daniel

AU - Griswold, Kettner

AU - Lin, Allen

AU - Zhao, Tongtong

AU - Marshall, Jamie L.

AU - Chen, Fei

PY - 2020/2/1

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

Abstract

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