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

被引：79

作者：

Chen, Haiqi ^{[1
]}

Liu, Sophia ^{[1
,2
,3
]}

Padula, Samuel ^{[1
]}

Lesman, Daniel ^{[1
]}

Griswold, Kettner ^{[4
,5
,6
,7
]}

Lin, Allen ^{[8
,9
]}

Zhao, Tongtong ^{[1
,10
]}

Marshall, Jamie L. ^{[1
]}

Chen, Fei ^{[1
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[2] Harvard Univ, Biophys Program, Boston, MA 02115 USA

[3] MIT, Harvard MIT Div Hlth Sci & Technol, Cambridge, MA 02139 USA

[4] MIT, MIT Media Lab, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[5] Harvard Univ, Wyss Inst Biol Inspired Engn, Boston, MA 02115 USA

[6] Harvard Med Sch, Dept Genet, Boston, MA 02115 USA

[7] Charles Stark Draper Lab, Cambridge, MA USA

[8] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA

[9] Ragon Inst MGH MIT & Harvard, Cambridge, MA USA

[10] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

来源：

NATURE BIOTECHNOLOGY | 2020年 / 38卷 / 02期

基金：

美国国家科学基金会;

关键词：

T7; RNA-POLYMERASE; SOMATIC HYPERMUTATION; DIRECTED EVOLUTION; GENOMIC DNA; EXPRESSION; PROMOTER; BACTERIAL; PLASMID; ASSAY; BASE;

D O I：

10.1038/s41587-019-0331-8

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：165 / +

页数：7

共 27 条

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[2]

BONNER G, 1994, J BIOL CHEM, V269, P25120

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