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

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Efficient, continuous mutagenesis in human cells using a pseudo-random DNA editor.
Nature Biotechnology ( IF 33.1 ) Pub Date : 2019-12-16 , DOI: 10.1038/s41587-019-0331-8
Haiqi Chen ₁ , Sophia Liu _{1,

2,

3} , Samuel Padula ₁ , Daniel Lesman ₁ , Kettner Griswold _{4,

5,

6,

7} , Allen Lin _{8,

9} , Tongtong Zhao _{1,

10} , Jamie L Marshall ₁ , Fei Chen ₁

Affiliation

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.

中文翻译：

使用伪随机DNA编辑器在人细胞中进行有效，连续的诱变。

在这里，我们描述了TRACE（T7聚合酶驱动的连续编辑），一种使用融合到T7 RNA聚合酶的胞苷脱氨酶在人细胞中进行连续，有针对性的诱变的方法。在整合到基因组中的T7启动子的控制下，TRACE在多个细胞世代中诱导了高速率的诱变。我们在MEK1抑制剂抗性筛选中使用了TRACE，并鉴定了功能相关的突变。

更新日期：2019-12-17

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