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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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